U.S. patent application number 10/863116 was filed with the patent office on 2005-10-06 for porcine cmp-n-acetylneuraminic acid hydroxylase gene.
This patent application is currently assigned to University of Pittsburgh. Invention is credited to Koike, Chihiro.
Application Number | 20050223418 10/863116 |
Document ID | / |
Family ID | 33511785 |
Filed Date | 2005-10-06 |
United States Patent
Application |
20050223418 |
Kind Code |
A1 |
Koike, Chihiro |
October 6, 2005 |
Porcine CMP-N-Acetylneuraminic acid hydroxylase gene
Abstract
The present invention provides porcine
CMP-N-Acetylneuraminic-Acid Hydroxylase (CMP-Neu5Ac hydroxylase)
protein, cDNA, and genomic DNA regulatory sequences. Furthermore,
the present invention includes porcine animals, tissues, and
organs, as well as cells and cell lines derived from such animals,
tissues, and organs, which lack expression of functional CMP-Neu5Ac
hydroxylase. Such animals, tissues, organs, and cells can be used
in research and in medical therapy, including in
xenotransplantation, and in industrial livestock farming
operations.
Inventors: |
Koike, Chihiro; (Pittsburgh,
PA) |
Correspondence
Address: |
KING & SPALDING LLP
191 PEACHTREE STREET, N.E.
ATLANTA
GA
30303-1763
US
|
Assignee: |
University of Pittsburgh
|
Family ID: |
33511785 |
Appl. No.: |
10/863116 |
Filed: |
June 7, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60476396 |
Jun 6, 2003 |
|
|
|
Current U.S.
Class: |
800/8 ; 435/191;
435/320.1; 435/325; 435/69.1; 536/23.2; 536/53 |
Current CPC
Class: |
A61P 43/00 20180101;
C12N 9/0071 20130101; A01K 67/0271 20130101; C12N 15/8509 20130101;
A61P 25/28 20180101; A61P 7/04 20180101; A01K 2227/108 20130101;
A01K 67/0276 20130101; A01K 2267/02 20130101; A01K 2267/025
20130101; A01K 2217/075 20130101 |
Class at
Publication: |
800/008 ;
536/023.2; 435/069.1; 435/191; 435/320.1; 435/325; 536/053 |
International
Class: |
A01K 067/00; C07H
021/04; C08B 037/00; C12N 009/06 |
Claims
We claim:
1. An isolated full length cDNA sequence encoding a porcine
CMP-N-Acetyneuraminic-Acid hydroxylase protein.
2. The cDNA of claim 1 wherein the sequence is SEQ ID NO 1.
3. The cDNA of claim 1 wherein the sequence is SEQ ID NO 3.
4. The cDNA of claim 1 wherein the sequence is SEQ ID NO 5.
5. The cDNA of claim 1 wherein the sequence is SEQ ID NO 7.
6. An isolated amino acid sequence encoding a
CMP-N-Acetyneuraminic-Acid hydroxylase protein wherein the sequence
is SEQ ID NO 2.
7. An isolated amino acid sequence encoding a
CMP-N-Acetyneuraminic-Acid hydroxylase protein wherein the sequence
is SEQ ID NO 4.
8. An isolated amino acid sequence encoding a
CMP-N-Acetyneuraminic-Acid hydroxylase protein wherein the sequence
is SEQ ID NO 6.
9. An isolated amino acid sequence encoding a
CMP-N-Acetyneuraminic-Acid hydroxylase protein wherein the sequence
is SEQ ID NO 8.
10. A nucleic acid construct comprising a full length cDNA sequence
encoding a porcine CMP-N-Acetyneuraminic-Acid hydroxylase
protein.
11. The construct of claim 10 wherein the cDNA is SEQ ID NO I.
12. The construct of claim 10 wherein the cDNA is SEQ ID NO 3.
13. The construct of claim 10 wherein the cDNA is SEQ ID NO 5.
14. The construct of claim 10 wherein the cDNA is SEQ ID NO 7.
15. The construct any one of claims 10-15 further comprising a
promoter.
16. The construct of any one of claims 10-15 further comprising a
selectable marker.
17. The construct of claim 16, wherein the selectable marker is
green fluorescent protein.
18. A transfected cell comprising the construct of any one of
claims 10-17.
19. A cell expressing a porcine CMP-N-Acetyneuraminic-Acid
hydroxylase protein.
20. The cell of claim 19 wherein the protein sequence is selected
from the group consisting of SEQ ID No 2, SEQ ID No 4, SEQ ID No 6
and SEQ ID No 8.
21. An isolated nucleotide sequence homologous to a nucleotide
sequence selected from the group consisting of SEQ ID No 1, SEQ ID
No 3, SEQ ID No 5 and SEQ ID No 7.
22. The nucleotide sequence of claim 21, wherein the sequence is at
least 80% homologous to the nucleotide sequence selected from the
group consisting of SEQ ID No 1, SEQ ID No 3, SEQ ID No 5 and SEQ
ID No 7.
23. The nucleotide sequence of claim 21, wherein the sequence is at
least 90% homologous to the nucleotide sequence selected from the
group consisting of SEQ ID No 1, SEQ ID No 3, SEQ ID No 5 and SEQ
ID No 7.
24. A nucleic acid construct comprising a nucleotide sequence
homologous to a nucleotide sequence selected from the group
consisting of SEQ ID No 1, SEQ ID No 3, SEQ ID No 5 and SEQ ID No
7.
25. An isolated nucleotide sequence that hybridizes to a nucleotide
sequence selected from the group consisting of SEQ ID No 1, SEQ ID
No 3, SEQ ID No 5 and SEQ ID No 7.
26. The nucleotide sequence of claim 25 that hybridizes under
stringent conditions.
27. A vector comprising a nucleotide sequence selected from the
group consisting of SEQ ID No 1, SEQ ID No 3, SEQ ID No 5 and SEQ
ID No 7.
28. A plasmid comprising a nucleotide sequence selected from the
group consisting of SEQ ID No 1, SEQ ID No 3, SEQ ID No 5 and SEQ
ID No 7.
29. An isolated nucleotide comprising a nucleotide sequence of SEQ
ID No 46.
30. An isolated nucleotide comprising a nucleotide sequence of SEQ
ID No 47.
31. An isolated nucleotide comprising a nucleotide sequence of SEQ
ID No 48.
32. An isolated nucleotide sequence selected from the group
consisting of SEQ ID No 10, SEQ ID No 1, SEQ ID No 12, SEQ ID No
13, SEQ ID No 14, SEQ ID No 15, SEQ ID No 16, SEQ ID No 17, SEQ ID
No 18, SEQ ID No 19, SEQ ID No 20, SEQ ID No 21, SEQ ID No 22, SEQ
ID No 22, SEQ ID No 23, SEQ ID No 24, SEQ ID No 25, SEQ ID No 26,
SEQ ID No 27 and SEQ ID No 28.
33. An isolated nucleotide sequence selected from the group
consisting of SEQ ID No 9, SEQ ID No 29, SEQ ID No 30, SEQ ID No
31, SEQ ID No 32, SEQ ID No 33, SEQ ID No 34, SEQ ID No 35, SEQ ID
No 36, SEQ ID No 37, SEQ ID No 38, SEQ ID No 39, SEQ ID No 40, SEQ
ID No 41, SEQ ID No 42, SEQ ID No 43, SEQ ID No 44 and SEQ ID No
45.
34. A vector comprising a sequence of any of claims 31-33.
35. A nucleic acid construct comprising a sequence of any of claims
31-33.
36. A plasmid comprising a sequence of any of claims 31-33.
37. A nucleic acid construct comprising at least 17 contiguous
nucleic acids of a sequence selected from the group comprising SEQ
ID No 9, SEQ ID No 10, Seq ID No 11, SEQ ID No 12, SEQ ID No 13,
SEQ ID No 19, SEQ ID No 21, SEQ ID No 25, SEQ ID No 29, SEQ ID No
30, SEQ ID No31, SEQ ID No 32, SEQ ID No 33, SEQ ID No 39, SEQ ID
No 40, SEQ ID No 41, SEQ ID No 42, SEQ ID No 43, SEQ ID No 44 and
SEQ ID No 45.
38. A nucleic acid construct comprising at least 17 contiguous
nucleic acids of SEQ ID No 47.
39. A nucleic acid construct comprising at least 2,775 contiguous
nucleic acids of SEQ ID Nos 48.
40. A nucleic acid construct comprising at least 1,750 contiguous
nucleic acids of SEQ ID Nos 49.
41. A nucleic acid construct comprising at least 150 contiguous
nucleic acids of SEQ ID Nos 50.
42. An isolated nucleotide comprising a sequence homologous to a
nucleotide sequence of any of claims 29 to 33.
43. The nucleotide of claim 41, wherein the sequence is at least
80% homologous to a nucleotide sequence of any of claims 29 to
33.
44. The nucleotide of claim 21, wherein the sequence is at least
90% homologous to a nucleotide sequence of any of claims 29 to
33.
45. An isolated nucleotide that hybridizes to a nucleotide sequence
of any of claims 29 to 33.
46. The nucleotide of claim 45 that hybridizes under stringent
conditions.
47. A targeting vector comprising: (a) a first nucleotide sequence
comprising at least 17 contiguous nucleic acids homologous to SEQ
ID No 47; (b) a selectable marker gene; and (c) a second nucleotide
sequence comprising at least 17 contiguous nucleic acids homologous
to SEQ ID No 48.
48. A targeting vector comprising: (a) a first nucleotide sequence
comprising at least 17 contiguous nucleic acids homologous to SEQ
ID No 48; (b) a selectable marker gene; and (c) a second nucleotide
sequence comprising at least 17 contiguous nucleic acids homologous
to SEQ ID No 48; wherein the first and second nucleotide sequences
do not overlap.
49. The targeting vector of claim 47 or 48 wherein the selectable
marker is green fluorescent protein.
50. The targeting vector of claim 47 or 48 wherein the first
nucleotide sequence represents the 5' recombination arm.
51. The targeting vector of claim 47 or 48 wherein the second
nucleotide sequence represents the 3' recombination arm.
52. The targeting vector of claim 49 wherein the selectable marker
is green fluorescent protein.
53. The targeting vector of claim 48 wherein the first nucleotide
sequence is homologous to a nucleotide sequence comprising at least
17 contiguous nucleotides of Intron 5.
54. The targeting vector of claim 48 wherein the second nucleotide
sequence is homologous to a nucleotide sequence comprising at least
17 contiguous nucleotides of Intron 6.
55. A cell transfected with the targeting vector of any one of
claims 47 to 54.
56. The cell of claim 55 wherein at least one allele of a porcine
CMP-N-Acetyeuraminic-Acid hydroxylase gene has been rendered
inactive via homologous recombination.
57. A porcine animal comprising the cell of claim 55.
58. The animal of claim 57 wherein at least one allele of a porcine
CMP-N-Acetyeuraminic-Acid hydroxylase gene has been rendered
inactive via homologous resombination.
59. An organ obtained from the animal of claim 58.
60. A tissue obtained from the animal of claim 58.
61. The organ of claim 59 wherein the organ is selected from the
group consisting of heart, lung, kidney and liver.
62. A method to produce a genetically modified cells comprising:
(a) transfecting a porcine cell with the targeting vector of claim
47 or 48; and (b) selecting a transfected cell in which at least
one allele of a porcine CMP-N-Acetyeuraminic-Acid hydroxylase gene
has been rendered inactive.
63. A method to produce a genetically modified animal comprising:
(a) transfecting a porcine cell with the targeting vector of claim
47 or 48; (b) selecting a tranfected cell in which at least one
allele of a porcine CMP-N-Acetyeuraminic-Acid hydroxylase gene has
been rendered inactive (a nuclear donor cell); (c) transferring the
nucleus of the nuclear donor cell into an enucleated oocyte to
produce an embryo; and (d) allowing the embryo to develop into an
animal.
64. An organ derived from the animal of claim 63.
65. The organ of claim 64 wherein the organ is selected from the
group consisting of heart, lung, kidney and liver.
66. A tissue derived from the animal of claim 63.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional patent
application Ser. No. 60/476,396, filed Jun. 6, 2003.
FIELD OF THE INVENTION
[0002] The present invention provides porcine
CMP-N-Acetylneuraminic-Acid Hydroxylase (CMP-Neu5Ac hydroxylase)
protein, cDNA, and genomic DNA regulatory sequences. Furthermore,
the present invention includes porcine animals, tissues, and
organs, as well as cells and cell lines derived from such animals,
tissues, and organs, which lack expression of functional CMP-Neu5Ac
hydroxylase. Such animals, tissues, organs, and cells can be used
in research and in medical therapy, including in
xenotransplantation, and in industrial livestock farming
operations. In addition, methods are provided to prepare organs,
tissues, and cells lacking the porcine CMP-Neu5Ac hydroxylase gene
for use in xenotranplantation.
BACKGROUND OF THE INVENTION
[0003] The unavailability of acceptable human donor organs, the low
rate of long term success due to host versus graft rejection, and
the serious risks of infection and cancer are the main challenges
now facing the field of tissue and organ transplantation. Because
the demand for acceptable organs exceeds the supply, many people
die each year while waiting for organs to become available. To help
meet this demand, research has been focused on developing
alternatives to allogenic transplantation. Dialysis is available to
patients suffering from kidney failure, artificial heart models
have been tested, and other mechanical systems have been developed
to assist or replace failing organs. Such approaches, however, are
quite expensive. The need for frequent and periodic access to
dialysis machines greatly limits the freedom and quality of life of
patients undergoing such therapy.
[0004] Xenograft transplantation represents a potentially
attractive alternative to artificial organs for human
transplantation. The potential pool of nonhuman organs is virtually
limitless. Pigs are considered the most likely source of xenograft
organs. The supply of pigs is plentiful, breeding programs are well
established, and their organ size and physiology are compatible
with humans. Therefore, xenotransplantation with pig organs offers
a potential solution to the shortage of organs available for
clinical transplantation.
[0005] Host rejection of such cross-species tissue remains a major
concern in this area. The immunological barriers to
xenotransplantation have been, and remain, formidable. The first
immunological hurdle is "hyperacute rejection" (HAR). HAR is
defined by the ubiquitous presence of high titers of pre-formed
natural antibodies binding to the foreign tissue. The binding of
these natural antibodies to target epitopes on the donor organ
endothelium is believed to be the initiating event in HAR. This
binding, within minutes of perfusion of the donor organ with the
recipient blood, is followed by complement activation, platelet and
fibrin deposition, and ultimately by interstitial edema and
hemorrhage in the donor organ, all of which cause failure of the
organ in the recipient (Strahan, et al. (1996) Frontiers in
Bioscience 1, pp. 34-41).
[0006] Some noted xenotransplants of organs from apes or old-world
monkeys (e.g., baboons) into humans have been tolerated for months
without rejection. However, such attempts have ultimately failed
due to a number of immunological factors. Even with heavy
immunosupression to suppress HAR, a low-grade innate immune
response, attributable in part to failure of complement regulatory
proteins (CRPs) within the graft tissue to control activation of
heterologous complement on graft endothelium, ultimately leads to
destruction of the transplanted organs (Starzl, Immunol. Rev., 141,
213-44 (1994)). In an effort to develop a pool of acceptable organs
for xenotransplantation into humans, researchers have engineered
animals that produce human CRPs, an approach which has been
demonstrated to delay, but not eliminate, xenograft destruction in
primates (McCurry, et al., Nat. Med., 1, 423-27 (1995); Bach et
al., Immunol. Today, 17, 379-84 (1996)).
[0007] In addition to complement-mediated attack, human rejection
of discordant xenografts appears to be mediated by a common
antigen: the galactose-.alpha.(1,3)-galactose (gal-.alpha.-gal)
terminal residue of many glycoproteins and glycolipids (Galili et
al., Proc. Nat. Acad. Sci., (USA), 84, 1369-73 (1987); Cooper, et
al., Immunol. Rev., 141, 31-58 (1994); Galili, et al., Springer
Sem. Immunopathol, 15, 155-171 (1993); Sandrin, et al., Transplant
Rev., 8, 134 (1994)). This antigen is chemically related to the
human A, B, and 0 blood antigens, and it is present on many
parasites and infectious agents, such as bacteria and viruses. Most
mammalian tissue also contains this antigen, with the notable
exception of old world monkeys, apes and humans. (see, Joziasse, et
al., J. Biol. Chem., 264, 14290-97 (1989). Individuals without such
carbohydrate epitopes produce abundant naturally occurring
antibodies (IgM as well as IgG) specific to the epitopes. Many
humans show significant levels of circulating IgG with specificity
for gal-.alpha.-gal carbohydrate determinants (Galili, et al., J.
Exp. Med., 162, 573-82 (1985); Galili, et al., Proc. Nat. Acad.
Sci. (TJSA), 84, 1369-73 (1987)). The .alpha.-galactosyltransferase
(.alpha.-GT) enzyme catalyzes the formation of gal-.alpha.-gal
moieties. Research has focused on the modulation or elimination of
this enzyme to reduce or eliminate the expression of
gal-.alpha.-gal moieties on the cell surface of xenotissue.
[0008] The elimination of the .alpha.-galactosyltransferase gene
from porcine has long been considered one of the most significant
hurdles to accomplishing xenotransplantation from pigs to humans.
Two alleles in the pig genome encode the .alpha.-GT gene. Single
allelic knockouts of the .alpha.-GT gene in pigs were reported in
2002 (Dai, et al. Nature Biotechnol., 20:251 (2002); Lai, et al.,
Science, 295:1089 (2002)).
[0009] Recently, double allelic knockouts of the .alpha.-GT gene
have been accomplished (Phelps, et al., Science, 299: pp. 411-414
(2003)). WO 2004/028243 to Revivicor Inc. describes porcine animal,
tissue, organ, cells and cell lines, which lack all expression of
functional .alpha.1,3 galactosyltransferase (.alpha.1,3-GT).
Accordingly, the animals, tissues, organs and cells lacking
functional expression of .alpha.1,3-GT can be used in
xenotransplantation and for other medical purposes.
[0010] PCT patent application WO 2004/016742 to Immerge
Biotherapeutics, Inc. describes .alpha.(1,3)-galactosyltransferase
null cells, methods of selecting GGTA-1 null cells,
.alpha.(1,3)-galactosyltransferase null swine produced therefrom
(referred to as a viable GGTA-1 null swine), methods for making
such swine, and methods of using cells, tissues and organs of such
a null swine for xenotransplantation.
[0011] One of the earliest known xenoantigens other than
gal-.alpha.-gal is an epitope that Hanganutiu Deicher antibodies
recognize, and which have long been associated with serum disease.
The epitope has been identified as N-glycolylneuraminic acid
(Neu5Gc), a member of the sialic acid family of carbohydrates.
Among carbohydrates, sialic acids are abundant and ubiquitous.
Sialic acid is a generic designation used for N-acylneuraminic
acids (Neu5Acyl) and their derivatives. N-Acetylneuraminic acid
(Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) are two of the most
abundant derivatives of sialic acids.
[0012] The Neu5Gc epitope is located in the terminal position in
the glycan chains of glycoconjugates. Due to this exposed position,
it plays an important role in cellular recognition, e.g. in the
case of inflammatory reactions, maturation of immune cells,
differentiation processes, hormone-, pathogen- and toxin binding
(Varki, A., Glycobiology, 2, pp. 25-40 (1992)).
[0013] Glycoconjugates containing Neu5Gc are immunogenic in humans.
In healthy humans, Neu5Gc is not detectable, although Neu5Gc is
abundant in most mammals. The lack of Neu5Gc in man is due to an
exon deletion in the human gene that prevents the formation of
functional enzyme (Chou, H. H., et al. Proc. Natl. Acad. Sci.
(USA), 95, pp. 11751-11756 (1998); Irie, A., et al. J. Biol. Chem.,
273, pp. 15866-15871 (1998)). Thus, Neu5Gc-containing
glycoconjugates act as antigens and can induce the formation of
antibodies. Historically, the antibodies have been referred to as
Hanganutziu-Deicher (HD) antigens and antibodies (Hanganutziu, M.,
CR Soc. Biol. (Paris), 91, p. 1457 (1924); Deicher, H., Z. Hyg.,
106, p. 561 (1926)). Hanganutziu-Deicher antigens are detectable in
many human tumors (colon carcinoma, retinoblastoma, melanoma and
carcinoma of the breast) as well as in chicken tumor tissues
(Higashi, H., et al. Cancer Res., 45, pp. 3796-3802 (1985)).
Although the amount of antigen in tumors is very small (usually
less than 1% of the total amount of sialic acid, often in the range
of from 0.01 to 0.1%), it is capable of inducing the formation of
Hanganutziu-Deicher antibodies (Higashihara, T., et al., Int Arch
Allergy Appl Immunol., 95, pp. 231-235 (1991)). This immunological
reaction is a potential barrier to xenotransplantation of
Neu5Gc-containing pig organs to humans.
[0014] The Neu5Gc epitope is formed by the addition of a hydroxyl
group to the N-acetyl moiety of Neu5Ac. The enzyme that catalyzes
the hydroxylation is CMP-Neu5Ac hydroxylase. Thus, the expression
of the CMP-Neu5Ac hydroxylase gene determines the presence of the
Neu5Gc epitope on cell surfaces. Purification studies of CMP-Neu5Ac
hydroxylase in mammals have shown that it is a soluble, cytosolic
oxygenase that is dependent on cytochrome b5 and cytochrome b5
reductase (Kawano, T., et al., J. Biol. Chem., 269, pp. 9024-9029
(1994); Schneckenburger, P., et al., Glycoconj. J., 11, pp. 194-203
(1994); Schlenzka, W., et al., Glycobiology, 4, pp. 675-683 (1994);
Kozutsumi, Y., et al., J. Biochem. (Tokyo), 108, pp. 704-706
(1990); and, Shaw, L., et al. Eur. J. Biochem., 219, pp. 1001-1011
(1994)).
[0015] Another important feature of Neu5Gc is that it acts as an
adhesion molecule for pathogens, allowing for entry into the cell
(Kelm, S. and Schauer, R., Int. Rev. Cytol, 179, pp. 137-240
(1997)). This causes disease and economic losses in certain
livestock species. Specifically, enterotoxigenic Escherichia coli
with K99 fimbriae infect newborn piglets by binding to Neu5Gc in
gangliosides such as Nue5Gca2.fwdarw.3Gal.beta.1.-
fwdarw.4Glc.beta.1.fwdarw.1' ceramide [GM3(Neu5Gc)],
N-glycolylsialoparagloboside and GM2(Neu5Gc) attached to intestinal
absorptive and mucus secreting cells, causing a potentially lethal
diarrhea (Malykh, Y., et. al., Biochem. J., 370, pp. 601-607
(2003); Kyogashima, M., et al., (1993); Teneberg, S., et al., FEBS
Letters, 263, pp. 10-14 (1990); Isobe, T., et al., Anal. Biochem.,
236, pp. 35-40 (1996); Lindahl, M. and Carlstedt, I., J. Gen.
Microbiol., 136, pp. 1609-1614 (1990); King, T. P., et al.,
Proceedings of the 6.sup.th International Symposium on Digestive
Physiology in Pigs, pp. 290-293, (1994)). Pig rotavirus infects pig
newborns causing diarrhea by binding to GM3(Neu5Gc). Pig
transmissible gastroenteritis coronavirus infects pigs via entry
into glycoconjugates containing .alpha.2,3-bound Neu5Gc (Schultz,
B., et al., J. Virol., 70, pp. 5634-5637 (1996)).
[0016] CMP-Neu5Ac hydroxylase has been isolated from mouse liver
and pig submandibular glands to homogeneity and characterized
(Kawano, T., et al., J. Biol. Chem., 269, pp. 9024-9029 (1994);
Schneckenburger, P., et al., Glycoconj. J., 11, pp. 194-203 (1994);
and, Schlenzka, W., et al., Glycobiology, 4, pp. 675-683
(1994)).
[0017] Schlenzka, et al. (Glycobiology, Vol. 4, pp. 675-683 (1994))
purified the enzyme from pig submandibular glands using ion
exchange chromatography, chromatography with immobilized triazin
dyes, hydrophobic interaction chromatography and gel filtration.
Schneckenburger et al. (Glycoconj. J., Vol. 11, pp. 194-203 (1994))
isolated the CMP-Neu5Ac hydroxylase from mouse liver. Both the
CMP-Neu5Ac hydroxylase from pig submandibular glands and the one
from mouse liver are soluble monomers having a molecular weight of
65 kDa. Their catalytic interactions with CMP-Neu5Ac and cytochrome
b5 are very similar to one another. The activity of these enzymes
seems to be dependent on an iron-containing prosthetic group.
[0018] JP-A 06 113838 describes the protein and DNA sequences of
murine CMP-Neu5Ac hydroxylase, as well as a monoclonal antibody
that specifically binds to the hydroxylase.
[0019] PCT Publication No. WO 97/03200A1 to Boehringer Manheim GMBH
discloses a partial cDNA for the porcine CMP-Neu5Ac hydroxylase.
This application discloses a cDNA sequence beginning in the middle
of Exon 8 of the CMP-Neu5Ac hydroxylase gene (further disclosed as
GenBank accession number Y15010).
[0020] Martensen, L., et al. (Eur. J. Biochem., Vol. 268, pp.
5157-5166 (2001)) discloses a full length amino acid sequence of
porcine CMP-Neu5Ac hydroxylase.
[0021] PCT Publication No. WO 02/088351 to RBC Biotechnology
discloses a partial cDNA and genomic sequence (exons 7-11 as well
as partial genomic sequence surrounding each exon) of porcine
CMP-NeuAc hydroxylase. In addition, methods are provided to
generate porcine cells and animals lacking the CMP-NeuAc hydoxylase
epitope, optionally, in combination with other genetic
modifications, such as inactivation of the
alpha-1,3-galactosyltransferase gene and/or insertion of complement
proteins.
[0022] It is an object of the present invention to provide genomic
and regulatory sequences of the porcine CMP-Neu5Ac hydroxylase
gene.
[0023] It is an object of the present invention to provide the full
length cDNA, as well as novel variants of the CMP-Neu5Ac
hydroxylase gene.
[0024] It is another object of the invention to provide novel
nucleic acid and amino acid sequences that encode the CMP-Neu5Ac
hydroxylase gene.
[0025] It is yet a further object of the present invention to
provide cells, tissues and/or organs deficient in the CMP-Neu5Ac
hydroxylase gene.
[0026] It is another object of the present invention to generate
animals, particularly pigs, lacking a functional CMP-Neu5Ac
hydroxylase gene.
[0027] It is yet a further object of the present invention to
provide cells, tissues and/or organs deficient in the CMP-Neu5Ac
hydroxylase gene for use in xenotransplantation of non-human organs
to human recipients in need thereof.
SUMMARY OF THE INVENTION
[0028] The full length cDNA sequence, peptide sequence, and genomic
organization of the porcine CMP-Neu5Ac hydroxylase gene has been
determined. To date, only partial cDNA and genomic sequences have
been identified. The present invention provides novel porcine
CMP-Neu5Ac hydroxylase protein, cDNA, cDNA variants, and genomic
DNA sequence. Furthermore, the present invention includes porcine
animals, tissues, and organs, as well as cells and cell lines
derived from such animals, tissue, and organs, which lack
expression of functional CMP-Neu5Ac hydroxylase. Such animals,
tissues, organs, and cells can be used in research and in medical
therapy, including xenotransplantation. In addition, methods are
provided to prepare organs, tissues, and cells lacking the porcine
CMP-Neu5Ac hydroxylase gene for use in xenotransplantation.
[0029] One aspect of the present invention provides the full length
cDNA of porcine CMP-Neu5Ac hydroxylase. The full length cDNA is
shown in Table 1 (SEQ ID No 1) and the full length peptide sequence
is provided in Table 2 (SEQ ID No 2). The start codon for the
full-length cDNA is located in the 3' portion of Exon 4, and the
stop codon is found in the 3' portion of Exon 17. Nucleotide and
amino acid sequences at least 80, 85, 90, 95, 98 or 99% homologous
to SEQ ID Nos 1 or 2 are provided. In addition, nucleotide and
peptide sequences that contain at least 10, 15, 17, 20, 25 or 30
nucleotide or amino acid sequences of SEQ ID Nos 1 or 2 are also
provided. Further provided is any nucleotide sequence that
hybridizes, optionally under stringent conditions, to SEQ ID No 1,
as well as, nucleotides homologous thereto.
[0030] In one embodiment, nucleic acid and peptide sequences
encoding three novel variants of CMP-Neu5Ac hydroxylase are
provided (Tables 3-8, FIG. 2). SEQ ID No 3 represents the cDNA of a
variant of the gene, variant-1, that includes Exons 1, 4, 5, 6, 7,
8, 9, 10, 11, 12, 13, 15a, 16, 17, and 18. SEQ ID No 5 represents
the cDNA of a variant of the gene, variant-2, that includes Exons
1, 4, 5, 6, 7, 8, 9, 10, 11, 12 and 12a. SEQ ID No 7 represents the
cDNA of a variant of the gene, variant-3, that includes Exons 1, 4,
5, 6, 7, 8, 9, 10, 11 and 11a. SEQ ID Nos 4, 6 and 8 represent the
amino acid sequences of variant-1, variant-2 and variant-3,
respectively. Nucleotide and amino acid sequences at least 80, 85,
90, 95, 98 or 99% homologous to SEQ ID Nos 3-8 are provided. In
addition, nucleotide and peptide sequences that contain at least
10, 15, 17, 20, 25, or 30 nucleotide or amino acid sequence of SEQ
ID Nos 3-8 are also provided. Further provided is any nucleotide
sequence that hybridizes, optionally under stringent conditions, to
SEQ ID Nos 3, 5 and 7, as well as, nucleotides homologous
thereto.
[0031] A further embodiment provides nucleic acid sequences
representing genomic DNA sequences of the CMP-Neu5Ac hydroxylase
gene (Table 9, FIG. 1). SEQ ID Nos 10-28 represent Exons 1, 4-11,
11a, 12, 12a, 13-15, 15a, 16-18, respectively, and SEQ ID Nos 29-45
represent Introns 1a, 1b, 4-15, 15a, 16, and 17, respectively. SEQ
ID No. 9 represents the 5' untranslated region of the CMP-Neu5Ac
hydroxylase gene. SEQ ID No. 46 (Table 10) represents the genomic
DNA and regulatory sequence of CMP-Neu5Ac hydroxylase.
[0032] In another embodiment, the genomic sequence of the porcine
CMP-Neu5Ac hydroxylase gene is represented by SEQ ID No. 47. SEQ ID
No. 47 represents the 5' contiguous genomic sequence containing 5'
UTR, Exon 1 and a portion of intronic sequence located 3' of Exon 1
(Table 11).
[0033] In another embodiment, the genomic sequence of the porcine
CMP-Neu5Ac hydroxylase gene is represented by SEQ ID No. 48. SEQ ID
NO. 48 represents a contiguous genomic sequence containing intronic
sequence located 5' to Exon 4, Exon 4, Intron 4, Exon 5, Intron 5,
Exon 6, Intron 6, Exon 7, Intron 7, Exon 8, Intron 8, Exon 9,
Intron 9, Exon 10, Intron 10, Exon 11, Intron 11, Exon 12, Intron
12, Exon 13, Intron 13, Exon 14, Intron 14, Exon 15, Intron 15,
Exon 16, Intron 16, Exon 17, Intron 17, and Exon 18 (Table 12). In
addition, nucleotide sequences that contain at least 2775, 2800,
2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900,
4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000,
9500 or 10,000 contiguous nucleotides of SEQ ID NO. 48 are
provided, as well as nucleotide sequences at least 80, 85, 90, 95,
98, or 99% homologous to SEQ ID NO. 48.
[0034] In another embodiment, the genomic sequence of the porcine
CMP-Neu5Ac hydroxylase gene is represented by SEQ ID No. 49. SEQ ID
NO. 49 represents contiguous genomic sequences containing Intronic
sequence 5' to Exon 4, Exon 4, Intron 4, Exon 5, Intron 5, Exon 6,
Intron 6, Exon 7, Intron 7 and Exon 8. Further, nucleotide
sequences that contain at least 1750, 1800, 1900, 2000, 2100, 2200,
2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300,
3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400,
4500, 4600, 4700, 4800, 4900, 5000, 5500, 6000, 6500, 7000, 7500,
8000, 8500, 9000, 9500, 10000, 11000, 12000, 13000, 14000, 15000,
16000, 17000, 18000, 19000, or 20000 contiguous nucleotides of SEQ
ID NO. 49 are provided, as well as nucleotide sequences at least
80, 85, 90, 95, 98, or 99% homologous to SEQ ID NO. 49.
[0035] In another embodiment, the genomic sequence of the porcine
CMP-Neu5Ac hydroxylase gene is represented by SEQ ID No. 50. SEQ ID
NO. 50 represents contiguous genomic sequences containing Exon 12,
Intron 12, Exon 13, Intron 13, Exon 14, Intron 14, Exon 15, Intron
15, Exon 16, Intron 16, Exon 17, Intron 17, and Exon 18 are
provided. Nucleotide sequences that contain at least 150, 200, 250,
300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900,
950, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500,
6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, 11000,
12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000 or 20,000
contiguous nucleotides of SEQ ID NO. 50 are provided, as well as
nucleotide sequences at least 80, 85, 90, 95, 98, or 99% homologous
to SEQ ID NO. 50.
[0036] In further embodiments, nucleotide and amino acid sequences
at least 80, 85, 90, 95, 98 or 99% homologous to SEQ ID Nos 9-45,
46, 47, 48, 49 and 50 are provided. In addition, nucleotide and
peptide sequences that contain at least 10, 15, 17, 20, 25, 30, 50,
100, 150, 200, 300, 400, 500 or 1000 contiguous nucleotide or amino
acid sequences of SEQ ID Nos 9-45, 46, 47, and 48 are also
provided. Further provided is any nucleotide sequence that
hybridizes, optionally under stringent conditions, to SEQ ID Nos
9-45, 46, 47, 48, 49 and 50, as well as, nucleotides homologous
thereto.
[0037] Another aspect of the present invention provides nucleic
acid constructs that contain cDNA or variants thereof encoding
CMP-Neu5Ac hydroxylase. These cDNA sequences can be derived from
Seq ID Nos. 1-8, or any fragment thereof. Constructs can contain
one, or more than one, internal ribosome entry site (IRES). The
construct can also contain a promoter operably linked to the
nucleic acid sequence encoding CMP-Neu5Ac hydroxylase, or,
alternatively, the construct can be promoterless. In another
embodiment, nucleic acid constructs are provided that contain
nucleic acid sequences that permit random or targeted insertion
into a host genome. In addition to the nucleic acid sequences the
expression vector can contain selectable marker sequences, such as,
for example, enhanced Green Fluorescent Protein (eGFP) gene
sequences, initiation and/or enhancer sequences, poly A-tail
sequences, and/or nucleic acid sequences that provide for the
expression of the construct in prokaryotic and/or eukaryotic host
cells.
[0038] In another embodiment, nucleic acid targeting vectors
constructs are also provided wherein homologous recombination in
somatic cells can be achieved. These targeting vectors can be
transformed into mammalian cells to target the CMP-Neu5Ac
hydroxylase gene via homologous recombination. In one embodiment,
the targeting vectors can contain a 3' recombination arm and a 5'
recombination arm that is homologous to the genomic sequence of a
CMP-Neu5Ac hydroxylase. The homologous DNA sequence can include at
least 15 bp, 20 bp, 25 bp, 50 bp, 100 bp, 500 bp, 1 kbp, 2 kbp, 4
kbp, 5 kbp, 10 kbp, 15 kbp, 20 kbp, or 50 kbp of sequence
homologous to the CMP-Neu5Ac hydroxylase sequence. In another
embodiment, the homologous DNA sequence can include one or more
intron and/or exon sequences. In a specific embodiment, the DNA
sequence can be homologous to Intron 5 and Intron 6 of the
CMP-Neu5Ac hydroxylase gene (see, for example, FIGS. 6-8). In
another specific embodiment, the DNA sequence can be homologous to
Intron 5, a 55 bp portion of Exon 6, and Intron 6 of the CMP-Neu5Ac
hydroxylase gene, and contain enhanced Green Fluorescent Protein
sequence in an in-frame orientation 3' to the 55 bp portion of Exon
6 (see, for example, FIGS. 10 and 11).
[0039] Another embodiment of the present invention provides
oligonucleotide primers capable of hybridizing to porcine
CMP-Neu5Ac hydroxylase cDNA or genomic sequence, such as Seq ID
Nos. 1, 3, 5, 7, 9-45, 46, 47 or 48. In a preferred embodiment, the
primers hybridize under stringent conditions to SEQ ID Nos. 1, 3,
5, 7, 9-45, 46, 47 or 48. Another embodiment provides
oligonucleotide probes capable of hybridizing to porcine CMP-Neu5Ac
hydroxylase nucleic acid sequences, such as SEQ ID Nos. 1, 3, 5, 7,
9-45, 46, 47, or 48. The polynucleotide primers or probes can have
at least 14 bases, 20 bases, preferably 30 bases, or 50 bases which
hybridize to a polynucleotide of the present invention. The probe
or primer can be at least 14 nucleotides in length, and in a
preferred embodiment, are at least 15, 20, 25, 28, or 30
nucleotides in length.
[0040] In another aspect of the present invention, mammalian cells
lacking at least one allele of the CMP-Neu5Ac hydroxylase gene
produced according to the process, sequences and/or constructs
described herein are provided. These cells can be obtained as a
result of homologous recombination. Particularly, by inactivating
at least one allele of the CMP-NeuAc hydroxylase gene, cells can be
produced which have reduced capability for expression of functional
Hanganutziu-Deicher antigens.
[0041] In embodiments of the present invention, alleles of the
CMP-Neu5Ac hydroxylase gene are rendered inactive according to the
process, sequences and/or constructs described herein, such that
the resultant CMP-Neu5Ac hydroxylase enzyme can no longer generate
Hanganutziu-Deicher antigens. In one embodiment, the CMP-Neu5Ac
hydroxylase gene can be transcribed into RNA, but not translated
into protein. In another embodiment, the CMP-Neu5Ac hydroxylase
gene can be transcribed in an inactive truncated form. Such a
truncated RNA may either not be translated or can be translated
into a nonfunctional protein. In an alternative embodiment, the
CMP-Neu5Ac hydroxylase gene can be inactivated in such a way that
no transcription of the gene occurs. In a further embodiment, the
CMP-Neu5Ac hydroxylase gene can be transcribed and then translated
into a nonfunctional protein.
[0042] In a further aspect of the present invention, porcine
animals are provided in which at least one allele of the CMP-Neu5Ac
hydroxylase gene is inactivated via a genetic targeting event
produced according to the process, sequences and/or constructs
described herein. In another aspect of the present invention,
porcine animals are provided in which both alleles of the
CMP-Neu5Ac hydroxylase gene are inactivated via a genetic targeting
event. The gene can be targeted via homologous recombination. In
other embodiments, the gene can be disrupted, i.e. a portion of the
genetic code can be altered, thereby affecting transcription and/or
translation of that segment of the gene. For example, disruption of
a gene can occur through substitution, deletion ("knock-out") or
insertion ("knock-in") techniques. Additional genes for a desired
protein or regulatory sequence that modulate transcription of an
existing sequence can be inserted.
[0043] In another aspect of the present invention, porcine cells
lacking one allele, optionally both alleles of the porcine
CMP-Neu5Ac hydroxylase gene can be used as donor cells for nuclear
transfer into enucleated oocytes to produce cloned, transgenic
animals. Alternatively, porcine CMP-Neu5Ac hydroxylase knockouts
can be created in embryonic stem cells, which are then used to
produce offspring. Offspring lacking a single allele of the
functional CMP-Neu5Ac hydroxylase gene produced according to the
process, sequences and/or constructs described herein can be breed
to further produce offspring lacking functionality in both alleles
through mendelian type inheritance. Cells, tissues and/or organs
can be harvested from these animals for use in xenotransplantation
strategies. The elimination of the Hanganutziu-Deicher antigens can
reduce the immune rejection of the transplanted cell, tissue or
organ due to the Neu5Gc epitope.
[0044] Alternatively, animals lacking at least one allele of the
CMP-Neu5Ac hydroxylase gene produced according to the process,
sequences and/or constructs described herein can be less
susceptible or resistant to enterotoxigenic infection and disease
such as, for example, E. Coli infection, rotavirus infection, and
gastroenteritis coronavirus. Such animals can be used, for example,
in commercial farming.
[0045] In one aspect of the present invention, a pig can be
prepared by a method in accordance with any aspect of the present
invention. Genetically modified pigs can be used as a source of
tissue and/or organs for transplantation therapy. A pig embryo
prepared in this manner or a cell line developed therefrom can also
be used in cell-transplantation therapy. Accordingly, there is
provided in a further aspect of the invention a method of therapy
comprising the administration of genetically modified cells lacking
porcine CMP-Neu5Ac hydroxylase to a patient, wherein the cells have
been prepared from an embryo or animal lacking CMP-Neu5Ac
hydroxylase. This aspect of the invention extends to the use of
such cells in medicine, e.g. cell-transplantation therapy, and also
to the use of cells derived from such embryos in the preparation of
a cell or tissue graft for transplantation. The cells can be
organized into tissues or organs, for example, heart, lung, liver,
kidney, pancreas, corneas, nervous (e.g. brain, central nervous
system, spinal cord), skin, or the cells can be islet cells, blood
cells (e.g. haemocytes, i.e. red blood cells, leucocytes) or
haematopoietic stem cells or other stem cells (e.g. bone
marrow).
[0046] In another aspect of the present invention, CMP-Neu5Ac
hydroxylase-deficient pigs also lack genes encoding other
xenoantigens, such as, for example, porcine iGb3 synthase (see, for
example, U.S. Patent Application No. 60/517,524), and/or porcine
Forssman synthase (see, for example, U.S. Patent Application No.
60/568,922). In another embodiment, porcine cells are provided that
lack the .alpha. 1,3 galactosyltransferase gene and the CMP-Neu5Ac
hydroxylase gene produced according to the process, sequences
and/or constructs described herein. In another embodiment, porcine
.alpha. 1,3 galactosyltransferase gene knockout cells are further
modified to knockout the CMP-Neu5Ac hydroxylase gene produced
according to the process, sequences and/or constructs described
herein. In addition, CMP-Neu5Ac hydroxylase deficient pigs produced
according to the process, sequences and/or constructs described
herein, optionally lacking one or more additional genes associated
with an adverse immune response, can be modified to express
complement inhibiting proteins, such as, for example, CD59, DAF,
and/or MCP can be further modified to eliminate the expression of
al least one allele of the CMP-Neu5Ac hydroxylase gene. These
animals can be used as a source of tissue and/or organs for
transplantation therapy. These animals can be used as a source of
tissue and/or organs for transplantation therapy. A pig embryo
prepared in this manner or a cell line developed therefrom can also
be used in cell-transplantation therapy.
DESCRIPTION OF THE INVENTION
[0047] Elimination of the CMP-Neu5Ac hydroxylase gene produced
according to the process, sequences and/or constructs described
herein can reduce a human beings immunological response to the
Neu5Gc epitope and remove an immunological barrier to
xenotransplantation. The present invention is directed to novel
nucleic acid sequences encoding the full-length cDNA and peptide.
Information about the genomic organization, intronic sequences and
regulatory regions of the gene are also provided. In one aspect,
the invention provides isolated and substantially purified cDNA
molecules having one of SEQ ID Nos: 1, 3, 5 or 7, or a fragment
thereof. In another aspect of the invention, DNA sequences
comprising the full-length genome of the CMP-NeuAc hydrolase gene
are provided in SEQ ID Nos 9-45, 46, 47, 48, 49 or 50 or fragments
thereof. In another aspect, primers for amplifying porcine
CMP-Neu5Ac hydroxylase cDNA or genomic sequence derived from SEQ ID
Nos. 1, 3, 5, 7, 9-45, 46, 47, 48, 49 or 50 are provided.
Additionally probes for identifying CMP-Neu5Ac hydroxylase nucleic
acid sequences derived from SEQ ID Nos. 1, 3, 5, 7, 9-45, 46, 47,
48, 49 or 50, or fragments thereof are provided. DNA represented by
SEQ ID Nos 9-45, 46, 47, 48, 49 or 50, or fragments thereof, can be
used to construct pigs lacking functional CMP-Neu5Ac hydroxylase
genes. Thus, the invention also provides a porcine chromosome
lacking a functional CMP-NeuAc hydroxylase gene and a transgenic
pig lacking a functional CMP-NeuAc hydroxylase protein produced
according to the process, sequences and/or constructs described
herein. Such pigs can be used as tissue sources for
xenotransplantation into humans. In an alternate embodiment,
CMP-NeuAc hydroxylase-deficient pigs produced according to the
process, sequences and/or constructs described herein also lack
other genes associated with adverse immune responses in
xenotransplantation, such as, for example, the .alpha.1,3
galactosyltransferase gene, iGb3 synthetase gene, or FSM synthase
gene. In another embodiment, pigs lacking CMP-Neu5Ac hydroxylase
produced according to the process, sequences and/or constructs
described herein and/or other genes associated with adverse immune
responses in xenotransplantation express complement inhibiting
factors such as, for example, CD59, DAF, and/or MCP.
BRIEF DESCRIPTION OF THE FIGURES
[0048] FIG. 1 represents the genomic organization of the porcine
CMP-Neu5Ac hydroxylase gene. Closed bars depict each numbered exon.
The length of the introns between the exons illustrates relative
distances. (Open boxes also represent exons that appear in some
variants (see FIG. 2); "start" and "stop" denote start and stop
codons, respectively) The approximate scale is depicted in the
bottom of the figure.
[0049] FIG. 2 depicts cDNA sequences of the CMP-Neu5Ac hydroxylase
gene. Variant-I contains exon 15a in place of exons 14 and 15.
Variant-2 contains exon 12a, and variant-3 contains exon 11a.
"Start" and "stop" denote the start and stop codons,
respectively.
[0050] FIG. 3 illustrates four non-limiting examples of targeting
vectors, along with their corresponding genomic organization. The
selectable marker gene in this particular non-limiting example is
eGFP (enhanced green fluorescent protein). eGFP can be inserted in
the DNA constructs to inactivate the porcine CMP-NeuAc hydroxylase
gene.
[0051] FIG. 4 illustrates transcription factor binding sites
located within exon 1 (228 bp) and its 5'-flanking region spanning
601 bp.
[0052] FIG. 5 depicts oligonucleotide sequences that can be used
for DNA construction of porcine CMP-Neu5Ac hydroxylase gene
targeting vector.
[0053] FIG. 6 is a schematic diagram illustrating the production of
a 3'-arm segment from the porcine CMP-Neu5Ac hydroxylase gene using
primers pDH3 and pDH4, and its insertion into a vector (pCRII).
[0054] FIG. 7 is a schematic diagram illustrating the production of
a 5'-arm segment from the porcine CMP-Neu5Ac hydroxylase gene using
primers pDH1 and pDH2, followed by pDH2a, pDH2b, and pDH2c, and its
insertion into a vector (pCRII) in which a 3'-arm has previously
been inserted.
[0055] FIG. 8 is a non-limiting example of a schematic illustrating
a targeting vector that can be utilized to delete Exon 6 of the
porcine CMP-Neu5Ac hydroxylase gene through homologous
recombination.
[0056] FIG. 9 represents oligonucleotide sequences used in
generating a enhanced green fluorescent protein expression vector
for use in a Knock-in strategy.
[0057] FIG. 10 is a schematic illustrating the insertion of a EGFP
fragment with a polyA signal into the targeting vector
pDH.DELTA.ex6.
[0058] FIG. 11 is a schematic illustrating a knock-in vector for
expression of eGFP.
[0059] FIG. 12 is a schematic illustrating homologous recombination
resulting in a frameshift between the targeting cassette DNA
construct (pDH.DELTA.ex6) and genomic DNA.
[0060] FIG. 13 is a schematic illustrating homologous recombination
resulting in a frameshift between the targeting cassette DNA
construct (pDH.DELTA.ex6) and genomic DNA.
Definitions
[0061] A "target DNA sequence" is a DNA sequence to be modified by
homologous recombination. The target DNA can be in any organelle of
the animal cell including the nucleus and mitochondria and can be
an intact gene, an exon or intron, a regulatory sequence or any
region between genes.
[0062] A "targeting DNA sequence" is a DNA sequence containing the
desired sequence modifications. The targeting DNA sequence can be
substantially isogenic with the target DNA.
[0063] A "homologous DNA sequence or homologous DNA" is a DNA
sequence that is at least about 80%, 85%, 90%, 95%, 98% or 99%
identical with a reference DNA sequence. A homologous sequence
hybridizes under stringent conditions to the target sequence,
stringent hybridization conditions include those that will allow
hybridization occur if there is at least 85% and preferably at
least 95% or 98% identity between the sequences.
[0064] An "isogenic or substantially isogenic DNA sequence" is a
DNA sequence that is identical to or nearly identical to a
reference DNA sequence. The term "substantially isogenic" refers to
DNA that is at least about 97-99% identical with the reference DNA
sequence, and preferably at least about 99.5-99.9% identical with
the reference DNA sequence, and in certain uses 100% identical with
the reference DNA sequence.
[0065] "Homologous recombination" refers to the process of DNA
recombination based on sequence homology.
[0066] "Gene targeting" refers to homologous recombination between
two DNA sequences, one of which is located on a chromosome and the
other of which is not.
[0067] "Non-homologous or random integration" refers to any process
by which DNA is integrated into the genome that does not involve
homologous recombination.
[0068] A "selectable marker gene" is a gene, the expression of
which allows cells containing the gene to be identified. A
selectable marker can be one that allows a cell to proliferate on a
medium that prevents or slows the growth of cells without the gene.
Examples include antibiotic resistance genes and genes which allow
an organism to grow on a selected metabolite. Alternatively, the
gene can facilitate visual screening of transformants by conferring
on cells a phenotype that is easily identified. Such an
identifiable phenotype can be, for example, the production of
luminescence or the production of a colored compound, or the
production of a detectable change in the medium surrounding the
cell.
[0069] The term "contiguous" is used herein in its standard
meaning, i.e., without interruption, or uninterrupted.
[0070] The term "porcine" refers to any pig species, including pig
species such as Large White, Landrace, Meishan, Minipig.
[0071] The term "oocyte" describes the mature animal ovum which is
the final product of oogenesis and also the precursor forms being
the oogonium, the primary oocyte and the secondary oocyte
respectively.
[0072] The term "fragment" means a portion or partial sequence of a
nucleotide or peptide sequence.
[0073] The terms "derivative" and "analog" means a nucleotide or
peptide sequence which retains essentially the same biological
function or activity as such nucleotide or peptide. For example, an
analog includes a proprotein which can be activated by cleavage of
the proprotein portion to produce an active mature polypeptide.
[0074] DNA (deoxyribonucleic acid) sequences provided herein are
represented by the bases adenine (A), thymine (T), cytosine (C),
and guanine(G).
[0075] Amino acid sequences provided herein are represented by the
following abbreviations:
1 A alanine P proline B aspartate or asparagine Q glutamine C
cysteine R arginine D aspartate S serine E glutamate T threonine F
phenylalanine G glycine V valine H histidine W tryptophan I
isoleucine Y tyrosine Z glutamate or glutamine K lysine L leucine M
methionine N asparagine
[0076] "Transfection" refers to the introduction of DNA into a host
cell. Cells do not naturally take up DNA. Thus, a variety of
technical "tricks" are utilized to facilitate gene transfer.
Numerous methods of transfection are known to the ordinarily
skilled artisan, for example, CaPO.sub.4 and electroporation. (J.
Sambrook, E. Fritsch, T. Maniatis, Molecular Cloning: A Laboratory
Manual, Cold Spring Laboratory Press, 1989). Transformation of the
host cell is the indicia of successful transfection.
I. Complete cDNA Sequence and Variants of the Porcine CMP-Neu5Ac
Hydroxylase Gene
[0077] One aspect of the present invention provides novel, full
length nucleic acid cDNA sequences of the porcine CMP-Neu5Ac
hydroxylase gene (FIG. 2, Table 1, Seq ID No 1). Another aspect of
the present invention provides predicted amino acid peptide
sequences of the porcine CMP-Neu5Ac hydroxylase gene (Table 2, Seq
ID No 2). The ATG start codon for the full-length cDNA is located
in the 3' portion of Exon 4, and the stop codon TAG is found in the
3' portion of Exon 17. Nucleic and amino acid sequences at least
90, 95, 98 or 99% homologous to Seq ID Nos 1 or 2 are provided. In
addition, nucleotide and peptide sequences that contain at least
10, 15, 17, 20 or 25 contiguous nucleic or amino acids of Seq ID
Nos 1 or 2 are also provided. Further provided are fragments,
derivatives and analogs of Seq ID Nos 1-2. Fragments of Seq ID Nos.
1-2 can include any contiguous nucleic acid or peptide sequence
that includes at least about 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55,
60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160,
170, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425,
450, 475, 500, 550, 600, 700, 750, 800, 850, 900, 1000, 5000 or
10,000 nucleotides.
2TABLE 1 Full length cDNA CCCGACGTCCTGGCAGCGCCCAGGCACTG Exons 1
& Seq ID No 1 TTATTGGTGCCTCCTGTGTCCACGCGCTT 4-18
CCCGGCCAGGCAGCCCTGGCGGATCCTAT TTTCTGTTCCCCCGATTCTGGTACCTC- TC
CCTCCCGCCCTCGGTGCGCAGCCGTCCTC CTGCAGTGCCTGCTCCTCCAGGGGCGAAA
CCGATCAGGGATCAGGCCACCCGCCTC- CT GAACATCCCTCCTTAGTTCCCACAGTCTA
ATGCCTTGTGGAAGCAAATGAGCCACAGA AGCTGAAGGAAAAACCACCATTCTTTC- TT
AATACCTGGAGAGAGGCAACGACAGACTA TGAGCAGCATCGAACAAACGACGGAGATC
CTGTTGTGCCTCTCACCTGCCGAAGCT- GC CAATCTCAAGGAAGGAATCAATTTTGTTC
GAAATAAGAGCACTGGCAAGGATTACATC TTATTTAAGAATAAGAGCCGCCTGAAG- GC
ATGTAAGAACATGTGCAAGCACCAAGGAG GCCTCTTCATTAAAGACATTGAGGATCTA
AATGGAAGGTCTGTTAAATGCACAAAA- CA CAACTGGAAGTTAGATGTAAGCAGCATGA
AGTATATCAATCCTCCTGGAAGCTTCTGT CAAGACGAACTGGTTGTAGAAAAGGAT- GA
AGAAAATGGAGTTTTGCTTCTAGAACTAA ATCCTCCTAACCCGTGGGATTCAGAACCC
AGATCTCCTGAAGATTTGGCATTTGGG- GA AGTGCAGATCACGTACCTTACTCACGCCT
GCATGGACCTCAAGCTGGGGGACAAGAGA ATGGTGTTCGACCCTTGGTTAATCGGT- CC
TGCTTTTGCGCGAGGATGGTGGTTACTAC ACGAGCCTCCATCTGATTGGCTGGAGAGG
CTGAGCCGCGCAGACTTAATTTACATC- AG TCACATGCACTCAGACCACCTGAGTTACC
CAACACTGAAGAAGCTTGCTGAGAGAAGA CCAGATGTTCCCATTTATGTTGGCAAC- AC
GGAAAGACCTGTATTTTGGAATCTGAATC AGAGTGGCGTCCAGTTGACTAATATCAAT
GTAGTGCCATTTGGAATATGGCAGCAG- GT AGACAAAAATCTTCGATTCATGATCTTGA
TGGATGGCGTTCATCCTGAGATGGACACT TGCATTATTGTGGAATACAAAGGTCAT- AA
AATACTCAATACAGTGGATTGCACCAGAC CCAATGGAGGAAGGCTGCCTATGAAGGTT
GCATTAATGATGAGTGATTTTGCTGGA- GG AGCTTCAGGCTTTCCAATGACTTTCAGTG
GTGGAAAATTTACTGAGGAATGGAAAGCC CAATTCATTAAAACAGAAAGGAAGAAA- CT
CCTGAACTACAAGGCTCGGCTGGTGAAGG ACCTACAACCCAGAATTTACTGCCCCTTT
CCTGGGTATTTCGTGGAATCCCACCCA- GC AGACAAGTATATTAAGGAAACAAACATCA
AAAATGACCCAAATGAACTCAACAATCTT ATCAAGAAGAATTCTGAGGTGGTAACC- TG
GACCCCAAGACCTGGAGCCACTCTTGATC TGGGTAGGATGCTAAAGGACCCAACAGAC
AGCAAGGGCATCGTAGAGCCTCCAGAA- GG GACTAAGATTTACAAGGATTCCTGGGATT
TTGGCCCATATTTGAATATCTTGAATGCT GCTATAGGAGATGAAATATTTCGTCAC- TC
ATCCTGGATAAAAGAATACTTCACTTGGG CTGGATTTAAGGATTATAACCTGGTGGTC
AGGATGATTGAGACAGATGAGGACTTC- AG CCCTTTGCCTGGAGGATATGACTATTTGG
TTGACTTTCTGGATTTATCCTTTCCAAAA GAAAGACCAAGCCGGGAACATCCATAT- GA
GGAAATTCGGAGCCGGGTTGATGTCATCA GACACGTGGTAAAGAATGGTCTGCTCTGG
GATGACTTGTACATAGGATTCCAAACC- CG GCTTCAGCGGGATCCTGATATATACCATC
ATCTGTTTTGGAATCATTTTCAAATAAAA CTCCCCCTCACACCACCTGACTGGAAG- TC
CTTCCTGATGTGCTCTGGGTAGAGAGGAC CTGAGCTGTCCCAGGGGTGCCCAACAACA
TGAAAAAATCAAGAATTTATTGCTGCT- AC GTCAAAGCTTATACCAGAGATTATGCCTT
ATAGACATTAGCAATGGATAATTATATGT TGCACTTGTGAAATGTGCACATATCCT- GT
TTATGAATCACCACATAGCCAGATTATCA ATATTTTACTTATTTCGTAAAAAATCCAC
AATTTTCCATAACAGAATCAACGTGTG- CA ATAGGAACAAGATTGCTATGGAAAACGAG
GGTAACAGGAGGAGATATTAATCCAAGCA TAGAAGAAATAGACAAATGAGGGGCCA- TA
AGGGGAATATAGGGAAGAGAAAAAAATTA AGATGGAATTTTAAAAGGAGAATGTAAAA
AATAGATATTTGTTCCTTAATAGGTTG- AT TCCTCAAATAGAGCCCATGAATATAATCA
AATAGGAAGGGTTCATGACTGTTTTCAAT TTTTCAAAAAGCTTTGTTGAAATCATA- GA
CTTGCAAAACAAGGCTGTAGAGGCCACCC TAAAATGGAAAATTTCACTGGGACTGAAA
TTATTTTGATTCAATGACAAAATTTGT- TA TTTACTGCGGATTATAAACTCTAACAAAT
AGCGATCTCTTTGCTTCATAAAAACATAA ACACTAGCTAGTAATAAAATGAGTTCT- GC
AG
[0078]
3TABLE 2 Full length Amino Acid Sequence M S S I E Q T T E I L L C
L S P A E A A Seq ID No 2 N L K E G I N F V R N K S T G K D Y I L F
K N K S R L K A C K N M C K H Q G G L F I K D I E D L N G R S V K C
T K H N W K L D V S S M K Y I N P P G S F C Q D E L V V E K D E E N
G V L L L E L N P P N P W D S E P R S P E D L A F G E V Q I T Y L T
H A C M D L K L G D K R M V F D P W L I G P A F A R G W W L L H E P
P S D W L E R L S R A D L I Y I S H M H S D H L S Y P T L K K L A E
R R P D V P I Y V G N T E R P V F W N L N Q S G V Q L T N I N V V P
F G I W Q Q V D K N L R F M I L M D G V H P E M D T C I I V E Y K G
H K I L N T V D C T R P N G G R L P M K V A L M M S D F A G G A S G
F P M T F S G G K F T E E W K A Q F I K T E R K K L L N Y K A R L V
K D L Q P R I Y C P F P G Y F V E S H P A D K Y I K E T N I K N D P
N E L N N L I K K N S E V V T W T P R P G A T L D L G R M L K D P T
D S K G I V E P P E G T K I Y K D S W D F G P Y L N I L N A A I G D
E I F R H S S W I K E Y F T W A G F K D Y N L V V R M I E T D E D F
S P L P G G Y D Y L V D F L D L S F P K E R P S R E H P Y E E I R S
R V D V I R H V V K N G L L W D D L Y I G F Q T R L Q R D P D I Y H
H L F W N H F Q I K L P L T P P D W K S F L M C S G
Variants
[0079] Another aspect of the present invention provides novel
nucleic acid cDNA sequences of three novel variants of CMP-Neu5Ac
hydroxylase gene transcript (FIG. 2, Tables 3, 5, and 7, Seq ID
Nos. 3, 5, and 7). Seq ID No 3 represents the cDNA of a variant of
the gene, variant-1, that includes Exons 1, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 15a, 16, 17, and 18. Exon 15a is a cryptic Exon that
normally appears in Intron 15, approxiametly 460 bp upstream of
Exon 16. The start codon for variant-I is located in Exon 4, while
the stop codon is located in Exon 17. Seq ID No 5 represents the
cDNA of a variant of the gene, variant-2, that includes Exons 1, 4,
5, 6, 7, 8, 9, 10, 11, 12 and 12a. Exon 12a is a cryptic Exon which
is retained from a partial sequence of Intron 12 (see SEQ ID. No.
21). The start codon for variant-2 is located in Exon 4, while the
stop condon is located in the terminal end of Exon 12a. Seq ID No 7
represents the cDNA of a variant of the gene, variant-3, that
includes Exons 1, 4, 5, 6, 7, 8, 9, 10, 11 and 11a. Exon 11a is a
cryptic Exon which is retained from a partial sequence of Intron 11
(see Seq ID No. 19). The start codon for variant-3 is located in
Exon 4, while the stop codon is located in Exon 11a. Another aspect
of the present invention provides predicted amino acid peptide
sequences of three novel variants of the porcine CMP-Neu5Ac
Hydroxylase gene transcript. Seq ID Nos 4, 6 and 8 represent the
amino acid sequences of variant-1, variant-2 and variant-3,
respectively. Nucleotide and amino acid sequences at least 80, 85,
90, 95, 98 or 99% homologous to Seq ID Nos 3-8 are provided. In
addition, nucleotide and peptide sequences that contain at least
10, 15, 17, 20, 25, 30, 50, 100, 150, 200, 300, 400, 500 or 1000
contiguous nucleotide or amino acid sequences of Seq ID Nos 3-8 are
also provided. Further provided are fragments, derivatives and
analogs of Seq ID Nos 3-8. Fragments of Seq ID Nos. 3-8 can include
any contiguous nucleic acid or peptide sequence that includes at
least about 10 bp, 15 bp, 17 bp, 20 bp, 50 bp, 100 bp, 500 bp, 1
kbp, 5 kbp or 10 kpb.
4TABLE 3 Variant-1 cDNA CCCGACGTCCTGGCAGCGCCCAGGCA Exons 1, 4- Seq
ID No 3 CTGTTATTGGTGCCTCCTGTGTCCAC 13, 15a, 16,
GCGCTTCCCGGCCAGGCAGCCCTGGC 17, 18 GGATCCTATTTTCTGTTCCCCCG- ATT
CTGGTACCTCTCCCTCCCGCCCTCGG TGCGCAGCCGTCCTCCTGCAGTGCCT
GCTCCTCCAGGGGCGAAACCGATCAG GGATCAGGCCACCCGCCTCCTGACAT
CCCTCCTTAGTTCCCACAGTCTAATG CCTTGTGGAAGCAAATGAGCCACAGA
AGCTGAAGGAAAAACCACCATTTCTT TCTTAATACCTGGAGAGAGGCAACGA
CAGACTATGAGCAGCATCGAACAAAC GACGGAGATCCTGTTGTGCCTCTCAC
CTGCCGAAGCTGCCAATCTCAAGGAA GGAATCAATTTTGTTCGAAATAAGAG
CACTGGCAAGGATTACATCTTATTTA AGAATAAGAGCCGCCTGAAGGCATGT
AAGAACATGTGCAAGCACCAAGGAGG CCTCTTCATTAAAGACATTGAGGATC
TAAATGGAAGGTCTGTTAAATGCACA AAACACAACTGGAAGTTAGATGTAAG
CAGCATGAAGTATATCAATCCTCCTG GAAGCTTCTGTCAAGACGAACTGGTT
GTAGAAAAGGATGAAGAAAATGGAGT TTTGCTTCTAGAACTAAATCCTCCTA
ACCCGTGGGATTCAGAACCCAGATCT CCTGAAGATTTGGCATTTCGGGGAAG
TGCAGATCACGTACCTTACTCACGCC TGCATGGACCTCAAGCTGGGGGACAA
GAGAATGGTGTTCGACCCTTGGTTAA TCGGTCCTGCTTTTGCGCGAGGATGG
TGGTTACTACACGAGCCTCCATCTGA TTGGCTGGAGAGGCTGAGCCGCGCAG
ACTTAATTTACATCAGTCACATGCAC TCAGACCACCTGAGTTACCCAACACT
GAAGAAGCTTGCTGAGAGAAGACCAG ATGTTCCCATTTATGTTGGCAACACG
GAAAGACCTGTATTTTGGAATCTGAA TCAGAGTGGCGTCCAGTTGACTAATA
TCAATGTAGTGCCATTGGAATATGGC AGCAGGTAGACAAAAATCTTCGATTC
ATGATCTTGATGGATGGCGTTCATCC TGAGATGGACACTTGCATTATTGTGG
AATACAAAGGTCATAAAATACTCAAT ACAGTGGATTGCACCAGACCCAATGG
AGGAAGGCTGCCTATGAAGGTTGCAT TAATGATGAGTGATTTTGCTGGAGGA
GCTTCAGGCTTTCCAATGACTTTCAG TGGTGGAAAATTTACTGAGGAATGGA
AAGCCCAATTCATTAAAACAGAAAGG AAGAAACTCCTGAACTACAAGGCTCG
GCTGGTGAAGGACCTACAACCCAGAA TTTACTGCCCCTTTCCTGGGTATTTC
GTGGAATCCCACCCAGCAGACAAGTA TATTAAGGAAACAAACATCAAAAATG
ACCCAAATGAACTCAACAATCTTATC AAGAAGAATTCTGAGGTGGTAACCTG
GACCCCAAGACCTGGAGCCACTCTTG ATCTGGGTAGGATGCTAAAGGACCCA
ACAGACAGATCCTGTGTCAGGAGTTG GGATTCTTTGAAGATTCGGAGCCGGG
TTGATGTCATCAGACACGTGGTAAAG AATGGTCTGCTCTGGGATGACTTGTA
CATAGGATTCCAAACCCGGCTTCAGC GGGATCCTGATATATACCATCATCTG
TTTTGGAATCATTTTCAAATAAAACT CCCCCTCACACCACCTGACTGGAAGT
CCTITCCTGATGTGCTCTGGGTAGAG AGGACCTGAGCTGTCCCAGGGGTGCC
CAACAACATGAAAAAATCAAGAATTT ATTGCTGCTACGTCAAAGCTTATACC
AGAGATTATGCCTTATAGACATTAGC AATGGATAATTATATGTTGCACTTGT
GAAATGTGCACATATCCTGTTTATGA ATCACCACATAGCCAGATTATCAATA
TTTTACTTATTTCGTAAAAAATCCAC AATTTTCCATAACAGAATCAACGTGT
GCAATAGGAACAAGATTGCTATGGAA AACGAGGGTAACAGGAGGAGATATTA
ATCCAAGCATAGAAGAAATAGACAAA TGAGGGGCCATAAGGGGAATATAGGG
AAGAGAAAAAAATTAAGATGGAATTT TAAAAGGAGAATGTAAAAAATAGATA
TTTGTTCCTTAATAGGTTGATTCCTC AAATAGAGCCCATGAATATAATCAAA
TAGGAAGGGTTCATGACTGTTTTCAA TTTTTCAAAAAGCTTTGTTGAAATCA
TAGACTTGCAAAACAAGGCTGTAGAG GCCACCCTAAAATGGAAAATTTCACT
GGGACTGAAATTATTTTGATTCAATG ACAAAATTTGTTATTTACTGCGGATT
ATAAACTCTAACAAATAGCGATCTCT TTGCTTCATAAAAACATAAACACTAG
CTAGTAATAAAATGAGTTCTGCAG
[0080]
5TABLE 4 Variant-1 Amino Acid Sequence M S S I E Q T T E I L L C L
S P A E A A Seq ID No 4 N L K E G I N F V R N K S T G K D Y I L F K
N K S R L K A C K N M C K H Q G G L F I K D I E D L N G R S V K C T
K H N W K L D V S S M K Y I N P P G S F C Q D E L V V E K D E E N G
V L L L E L N P P N P W D S E P R S P E D L A F G E V Q I T Y L T H
A C M D L K L G D K R M V F D P W L I G P A F A R G W W L L H E P P
S D W L E R L S R A D L I Y I S H M H S D H L S Y P T L K K L A E R
R P D V P I Y V G N T E R P V F W N L N Q S G V Q L T N I N V V P F
G I W Q Q V D K N L R F M I L M D G V H P E M D T C I I V E Y K G H
K I L N T V D C T R P N G G R L P M K V A L M M S D F A G G A S G F
P M T F S G G K F T E E W K A Q F I K T E R K K L L N Y K A R L V K
D L Q P R I Y C P F P G Y F V E S H P A D K Y I K E T N I K N D P N
E L N N L I K K N S E V V T W T P R P G A T L D L G R M L K D P T D
R S C V R S W D S L K I R S R V D V I R H V V K N G L L W D D L Y I
G F Q T R L Q R D P D I Y H H L F W N H F Q I K L P L T P P D W K S
F L M C S G
[0081]
6TABLE 5 Variant-2 cDNA CCCGACGTCCTGGCAGCGCCCAGGCAC Exons 1, 4- Seq
ID No 5 TGTTATTGGTGCCTCCTGTGTCCACGC 12, 12a
GCTTCCCGGCCAGGCAGCCCTGGCGGA TCCTATTTTCTGTTCCCCCGATTCTGG
TACCTCTCCCTCCCGCCCTCGGTGCGC AGCCGTCCTCCTGCAGTGCCTGCTCCT
CCAGGGGCGAAACCGATCAGGGATCAG GCCACCCGCCTCCTGAACATCCCTCCT
TAGTTCCCACAGTCTAATGCCTTGTGG AAGCAAATGAGCCACAGAAGCTGAAGG
AAAAACCACCATTCTTTCTTAATACCT GGAGAGAGGCAACGACAGACTATGAGC
AGCATCGAACAAACGACGGAGATCCTG TTGTGCCTCTCACCTGCCGAAGCTGCC
AATCTCAAGGAAGGAATCAATTTTGTT CGAAATAAGAGCACTGGCAAGGATTAC
ATCTTATTTAAGAATAAGAGCCGCCTG AAGGCATGTAAGAACATGTGCAAGCAC
CAAGGAGGCCTCTTCATTAAAGACATT GAGGATCTAAATGGAAGGTCTGTTAAA
TGCACAAAACACAACTGGAAGTTAGAT GTAAGCAGCATGAAGTATATCAATCCT
CCTGGAAGCTTCTGTCAAGACGAACTG GTTGTAGAAAAGGATGAAGAAAATGGA
GTTTTGCTTCTAGAACTAAATCCTCCT AACCCGTGGGATTCAGAACCCAGATCT
CCTGAAGATTTGGCATTTGGGGAAGTG CAGATCACGTACCTTACTCACGCCTGC
ATGGACCTCAAGCTGGGGGACAAGAGA ATGGTGTTCGACCCTTGGTTAATCGGT
CCTGCTTTTGCGCGAGGATGGTGGTTA CTACACGAGCCTCCATCTGATTGGCTG
GAGAGGCTGAGCCGCGCAGACTTAATT TACATCAGTCACATGCACTCAGACCAC
CTGAGTTACCCAACACTGAAGAAGCTT GCTGAGAGAAGACCAGATGTTCCCATT
TATGTTGGCAACACGGAAAGACCTGTA TTTTGGAATCTGAATCAGAGTGGCGTC
CAGTTGACTAATATCAATGTAGTGCCA TTTGGAATATGGCAGCAGGTAGACAAA
AATCTTCGATTCATGATCTTGATGGAT GGCGTTCATCCTGAGATGGACACTTGC
ATTATTGTGGAATACAAAGGTCATAAA ATACTCAATACAGTGGATTGCACCAGA
CCCAATGGAGGAAGGCTGCCTATGAAG GTTGCATTAATGATGAGTGATTTTGCT
GGAGGAGCTTCAGGCTTTCCAATGACT TTCAGTGGTGGAAAATTTACTGAGGAA
TGGAAAGCCCAATTCATTAAAACAGAA AGGAAGAAACTCCTGAACTACAAGGCT
CGGCTGGTGAAGGACCTACAACCCAGA ATTTACTGCCCCTTTCCTGGGTATTTC
GTGGAATCCCACCCAGCAGACAAGTAT GGCTGGATATGAGTGATTTAGCTGTGA
AACAACATGAAATGAGAAAGAATGATT AGTAGGGGTCTGGAGCTTATTTTAACA
AGCAGCCTGAAAACAGAAAGTATGAAT AAAAAAAATTAAATGCAAAAAAAAAAA
AAAAAAAAAAAAAAAAA
[0082]
7TABLE 6 Variant-2 Amino Acid Sequence M S S I E Q T T E I L L C L
S P A E A A Seq ID No 6 N L K E G I N F V R N K S T G K D Y I L F K
N K S R L K A C K N M C K H Q G G L F I K D I E D L N G R S V K C T
K H N W K L D V S S M K Y I N P P G S F C Q D E L V V E K D E E N G
V L L L E L N P P N P W D S E P R S P E D L A F G E V Q I T Y L T H
A C M D L K L G D K R M V F D P W L I G P A F A R G W W L L H E P P
S D W L E R L S R A D L I Y I S H M H S D H L S Y P T L K K L A E R
R P D V P I Y V G N T E R P V F W N L N Q S G V Q L T N I N V V P F
G I W Q Q V D K N L R F M I L M D G V H P E M D T C I I V E Y K G H
K I L N T V D C T R P N G G R L P M K V A L M M S D F A G G A S G F
P M T F S G G K F T E E W K A Q F I K T E R K K L L N Y K A R L V K
D L Q P R I Y C P F P G Y F V E S H P A D K Y G W I F Y I T C L R I
S
[0083]
8TABLE 7 Variant-3 cDNA CCCGACGTCCTGGCAGCGCCCAGGCAC Exons 1, 4- Seq
ID No 7 TGTTATTGGTGCCTCCTGTGTCCACGC 11, 11a
GCTTCCCGGCCAGGCAGCCCTGGCGGA TCCTATTTTCTGTTCCCCCGATTCTGG
TACCTCTCCCTCCCGCCCTCGGTGCGC AGCCGTCCTCCTGCAGTGCCTGCTCCT
CCAGGGGCGAAACCGATCAGGGATCAG GCCACCCGCCTCCTGAACATCCCTCCT
TAGTTCCCACAGTCTAATGCCTTGTGG AAGCAAATGAGCCACAGAAGCTGAAGG
AAAAACCACCATTCTTTCTTAATACCT GGAGAGAGGCAACGACAGACTATGAGC
AGCATCGAACAAACGACGGAGATCCTG TTGTGCCTCTCACCTGCCGAAGCTGCC
AATCTCAAGGAAGGAATCAATTTTGTT CGAAATAAGAGCACTGGCAAGGATTAC
ATCTTATTTAAGAATAAGAGCCGCCTG AAGGCATGTAAGAACATGTGCAAGCAC
CAAGGAGGCCTCTTCATTAAAGACATT GAGGATCTAAATGGAAGGTCTGTTAAA
TGCACAAAACACAACTGGAAGTTAGAT GTAAGCAGCATGAAGTATATCAATCCT
CCTGGAAGCTTCTGTCAAGACGAACTG GTTGTAGAAAAGGATGAAGAAAATGGA
GTTTTGCTTCTAGAACTAAATCCTCCT AACCCGTGTAGATGTAAGCAGCATGAA
GTATATCAATCCTCCTGGAAGGAAGTG CAGATCACGTACCTTACTCACGCCTGC
ATGGACCTCAAGCTGGGGGACAAGAGA ATGGTGTTCGACCCTTGGTTAATCGGT
CCTGCTTTTGCGCGAGGATGGTGGTTA CTACACGAGCCTCCATCTGATTGGCTG
GAGAGGCTGAGCCGCGCAGACTTAATT TACATCAGTCACATGCACTCAGACCAC
CTGAGTTACCCAACACTGAAGAAGCTT GCTGAGAGAAGACCAGATGTTCCCATT
TATGTTGGCAACACGGAAAGACCTGTA TTTTGGAATCTGAATCAGAGTGGCGTC
CAGTTGACTAATATCAATGTAGTGCCA TTTGGAATATGGCAGCAGGTAGACAAA
AATCTTCGATTCATGATCTTGATGGAT GGCGTTCATCCTGAGATGTCAATACAG
TGGATTGCACCAGACCCAATGGAGGAA GGCTGCCTATGAAGGTTGCATTAATGA
TGAGTGATTTTGCTGGAGGAGCTTCAG GCTTTCCAATGACTTTCAGTGGTGGAA
AATTTACTGGTAATTCTTTATATCAAA ATGATGCCAAGGAGTTGGCATGGCACT
TTGCTAAATGCTGTGTGAATCAATACA AAGATAATTAGGACATGGTTCTTCCTC
ACAAGAGGTGTGCAATCTTATTGGGAA ATCATACTTGCAAGTCACAAATATAGA
CTAAAGTTTCCAGCTGAGAATATGCTG ATGGAGCATGAAACACTAAGGAGACAG
GGAGAATCTCAGGAAAAATCAAGAATA ATTTGGATCAAATGGATTCCTGACATA
GAACATAGAGCTGATCAGAAAGAGTCT GACATTGGTAATCCAGGCTTAAGTGCT
CTTTGTATGTGGTTCAGAACAGAGTGT GGGCAGCCTGAGGGGGATACATACCCT
TGACCTCGTGGAAAGCTCATACGGGGG AGGGATGAGGCTAAGGAAGCCCCTCTA
AAGTGTGGGATTACGAGAGGTTGGGGG GGTGGTAGGGAAAATAGTGGTCAAAGA
GTATAAACTTCCAGTTACAAGATGAAT AAATTCTAGGGGTATAATAACAGCATG
GCACTATAGATAGCATATTGTACTATA TACTGGAAGTGCTGAGAGTAGATCTTA
CATGTTCTAACCACACACACACACACA CACACACACACACCACACACACACACC
ACACACACACACGTGCACACAAACAGA AATGGTAATTATGTGAGGTGATGGCGG
TGTTAATAACTTTATTGTGGTCATCAT TTAGCCATACATGCATGTCATGAAATC
ACCATGTTGTACACCTTAAAGTTATGT AATACTAGATGTCAGTTATATCTCAAA
GCTAGAAAAAATGTGGGGACCAAGGCA GAAGCTCTTCTGCTCTGTGTCTAAGGG
TGGTTCTGGGGCTGGGATGGGGAGGAT GGTTAAGTGGTATATTTTTTTCATACC
TTTGCTCAGTACTATCATTGTAAGTGT TCAATATATGTCTGCTTAATAAATTAA
TGTTTTTAGTAAAAAAAAAAAAAAAAA AAAAAAAAAAA
[0084]
9TABLE 8 Variant-3 Amino Acid Sequence M S S T E Q T T E I L L C L
S P A E A A Seq ID No 8 N L K E G I N F V R N K S T G K D Y I L F K
N K S R L K A C K N M C K H Q G G L F I K D I E D L N G R S V K C T
K H N W K L D V S S M K Y I N P P G S F C Q D E L V V E K D E E N G
V L L L E L N P P N P W D S E P R S P E D L A F G E V Q I T Y L T H
A C M D L K L G D K R M V F D P W L I G P A F A R G W W L L H E P P
S D W L E R L S R A D L I Y I S H M H S D H L S Y P T L K K L A E R
R P D V P I Y V G N T E R P V F W N L N Q S G V Q L T N I N V V P F
G I W Q Q V D K N L R F M I L M D G V H P E M D T C I I V E Y K G H
K I L N T V D C T R P N G G R L P M K V A L M M S D F A G G A S G F
P M T F S G G K F T G N S L Y Q N D A K E L A W H F A K C C V N Q Y
K D N
[0085] In other aspects of the present invention, nucleic acid
constructs are provided that contain cDNA or variants thereof
encoding CMP-Neu5Ac hydroxylase. These cDNA sequences can be SEQ ID
NO 1, 3, 5 or 7, or derived from SEQ ID Nos. 2, 4, 6, or 8 or any
fragment thereof. Constructs can contain one, or more than one,
internal ribosome entry site (IRES). The construct can also contain
a promoter operably linked to the nucleic acid sequence encoding
CMP-Neu5Ac hydroxylase, or, alternatively, the construct can be
promoterless. In another embodiment, nucleic acid constructs are
provided that contain nucleic acid sequences that permit random or
targeted insertion into a host genome. In addition to the nucleic
acid sequences the expression vector can contain selectable marker
sequences, such as, for example, enhanced Green Fluorescent Protein
(eGFP) gene sequences, initiation and/or enhancer sequences, poly
A-tail sequences, and/or nucleic acid sequences that provide for
the expression of the construct in prokaryotic and/or eukaryotic
host cells. Suitable vectors and selectable markers are described
below. The expression constructs can further contain sites for
transcription initiation, termination, and/or ribosome binding
sites. The constructs can be expressed in any prokaryotic or
eukaryotic cell, including, but not limited to yeast cells,
bacterial cells, such as E. Coli, mammalian cells, such as CHO
cells, and/or plant cells.
[0086] Promoters for use in such constructs, include, but are not
limited to, the phage lambda PL promoter, E. coli lac, E. coli trp,
E. coli phoA, E. coli tac promoters, SV40 early, SV40 late,
retroviral LTRs, PGKI, GALI, GALIO genes, CYCI, PH05, TRPI, ADHI,
ADH2, forglymaldehyde phosphate dehydrogenase, hexokinase, pyruvate
decarboxylase, phosphofructokinase, triose phosphate isomerase,
phosphoglucose isomerase, glucokinase alpha-mating factor
pheromone, PRBI, GUT2, GPDI promoter, metallothionein promoter,
and/or mammalian viral promoters, such as those derived from
adenovirus and vaccinia virus. Other promoters will be known to one
skilled in the art.
II. Genomic Sequences of the CMP-Neu5Ac Hydroxylase Gene
[0087] Nucleic acid sequences representing the genomic DNA
organization of the CMP-Neu5Ac hydroxylase gene (FIG. 1, Table 9)
are also provided. Seq ID Nos 10-28 represent Exons 1, 4-11, 11a,
12, 12a, 13-15, 15a, and 16-18, respectively. Exons 11a, 12a, and
15a are cryptic Exons that are retained in certain variant
transcripts of CMP-Neu5Ac hydroxylase. SEQ ID Nos 29-45 represent
Intronic sequence between Exon 1 and Exon 4 (hereinafter Intron 1a
and Intron 1b, respectively), 4-15, 15a, 16, and 17, respectively.
Intron 15a is the 3' downstream portion of Intron 15 that follows
the cryptic Exon 15a. Seq ID No. 9 represents the 5' untranslated
region of the porcine CMP-Neu5Ac hydroxylase gene. Nucleic acid
sequence representing the genomic DNA sequence of the porcine
CMP-Neu5Ac hydroxylase gene (Table 10, SEQ ID No. 46) is also
provided. In addition, contiguous genomic sequence representing the
5' contiguous genomic sequence containing 5' UTR, Exon 1 and a
portion of intronic sequence located between Exon 1 and Exon 4
(Intron 1a) (SEQ ID No. 47, Table 11) is provided. Contiguous
genomic sequence containing an intronic sequence located between
Exon 1 and Exon 4 (Intron 1b) through Exon 18 (SEQ ID No. 48, Table
12) is also provided. Nucleotide and amino acid sequences at least
80, 85, 90, 95, 98 or 99% homologous to SEQ ID Nos 9-45, 46, 47,
48, 49 and 50 are provided. In addition, nucleotide and peptide
sequences that contain at least 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55,
60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160,
170, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425,
450, 475, 500, 550, 600, 700, 750, 800, 850, 900, 1000, 5000 or
10,000 contiguous nucleotide or amino acid sequences of SEQ ID Nos
9-45, 46, 47, 48, 49 and 50 are also provided, as well as any
nucleotide sequence 80, 85, 90, 95, 98 or 99% homologous thereto.
Further provided are fragments, derivatives and analogs of SEQ ID
Nos 9-45, 46, 47, 48, 49, and 50. Fragments of Seq ID Nos. 9-45,
46, 47, 48, 49, and 50 can include any contiguous nucleic acid or
peptide sequence or at least about 10 bp, 15 bp, 17 bp, 20 bp, 50
bp, 100 bp, 500 bp, 1 kbp, 5 kbp or 10 kpb.
[0088] In addition, regulatory regions in the form of putative
transcription factor binding sites of the genomic sequence have
been identified (see FIG. 4). These binding sites are located in
the 5'UTR and Exon 1 of the porcine CMP-Neu5Ac hydroxylase genome,
and include binding sites for transcription factors such as, for
example, ETSF, MZFI, SF1, CMYB, MEF2, TATA, MEF2, NMP4, CAAT, API,
BRN2, SATBI, ATF, GATI, USF, WHN, NMP4, ZF5, NFKB, ZBP89, MOK2,
ZF5, NFY, and MYCMAX.
10TABLE 9 Genomic Organizational Sequences
ctgccagcctaagccacagccacagcaac 5'UTR Seq ID No 9
gctgggtctgagccatgtctgcagcctat gccagagctccccgcagcgccggatgctt
aacccactgagcaaggccagggattgaac cctcgtcctcatggatagcagttgagttg
tttccacggaactcttaggggaactcctg attattttttatttaaatttatatttctc
tgactttttcgtgtgctcatcagccactg actgtgtatctccattagtcatggtttgt
taactctgtcattcaaaccctcttcatcc ttgctacgcagataacatcattataataa
aatcgtgcctgaagaccagtgacgccccc aagctaagttactgcttcccctgggggga
aaaagaagcaccgcgcgggcgctgacacg aagtccgggcagaggaagacggggcagag
gaagacgggggagcagtgggagcagcggg cagggcgcgggaagcactggggatgttcc
gcgttggcaggagggtgttgggcgagctc ccggtgatgcaggggggaggagccttttc
cgaagtagcgggacaagagccacgggaag gaactgttctgagttcccagt
CCCGACGTCCTGGCAGCGCCCAGGCACTG Exon 1 Seq ID No 10
TTATTGGTGCCTCCTGTGTCCACGCGCTT CCCGGCCAGGCAGCCCTGGCGGATCCTAT
TTTCTGTTCCCCCGATTCTGGTACCTCTC CCTCCCGCCCTCGGTGCGCAGCCGTCCTC
CTGCAGTGCCTGCTCCTCCAGGGGCGAAA CCGATCAGGGATCAGGCCACCCGCCTCCT
GAACATCCCTCCTTAGTTCCCACAG gtgagaaggcttcgccgctgctgccgctg Intron Seq
ID No 29 gcgccggcagcgccctccacgcacttcgt 1a
agtgggcgcgcgccctcctgcattgtttc taaaagatttttttttatccgcttatgct
atcagttactgaggaagtatttacaaatc tactattattttgaatttgcctttttctc
cttatagtttatcagtatctcttgagact gttattggtgcctgcaaatttaaaatgat
tggggttttatgaggaagtgaacctttta tctttatgaaacgcctaactgaggcaatg
ttaattgcttaaaatactttctttattat cagtgtggccatgccagtgtcctcttggt
tagaatttgcctgat ctgccaaagctgggagatgggggaaagta Intron Seq ID No 30
gagtgggttattgaaactgaatatagagt 1b tcagcatctaaaagcgaggtagtagagga
ggaagctgtgtcaacggaaatactgagct gggttcacatcctctttctccacacag
TCTAATGCCTTGTGGAAGCAAATGAGCCA Exon 4 Seq ID No 11
CAGAAGCTGAAGGAAAAACCACCATTCTT TCTTAATACCTGGAGAGAGGCAACGACAG
ACTATGAGCAG gcaagtgagagggggctttagctgtcagg Intron 4 Seq ID No 31
gaaggcggagataaacccttgatgggtag gatggccattgaaaggaggggagaaattt
gccccagcaggtagccaccaagcttgggg acttggagggagggctttcaaacgtattt
tcataaaaaagacctgtggagctgtcaat gctcagggattctctcttaaaatctaaca
gtattaatctgctaaaacatttgcctttt catag CATCGAACAAACGACGGAGATCCTGTTG
Exon 5 Seq ID No 12 TGCCTCTCACCTGCCGAAGCTGCCAATC
TCAAGGAAGGAATCAATTTTGTTCGAAA TAAGAGCACTGGCAAGGATTACATCTTA
TTTAAGAATAAGAGCCGCCTGAAGGCAT GTAAGAACATGTGCAAGCACCAAGGAGG
CCTCTTCATTAAAGACATTGAGGATCTA AATGGAAG gtactgagaatcctttgctttctccctg
Intron 5 Seq ID No 32 gcgatcctttctcccaattaggtttggc
aggaaatgtgctcattgagaaatttta- a atgatccaatcaacatgctatttccccc
agcacatgcctaactttttcttaagctc ctttacggcagctctctgattttgattt
atgaccttgacttaatttcccatcctct ctgaagaactattgtttaaaatgtattc
ctagttgataaacagtgaaacttctaag gcacatgtgtgtgtgtgtgtgtgtgtgt
gtgtgtttaccagcttttatattcaaag actcaagcctcttttggatttcctttcc
tgctctctcagaagtgtgtgtgtgaggt gagtgcttgtccaaacactgccctagaa
cagagagactttccctgatgaaaacccg aaaaatggcagagctctagctgcacctg
gcctcaacagcggctcttctgatcattt cttggaagaacgagtgctggtacccctt
ttccccagccccttgattaaacctgcat atcgcttgcctccccatctcaggagcaa
ttctaggagggagggtgggctttctttt caggattgacaaagctacccagcttgca
aaccagggggatctggggggggggtttg cacctgatgctcccccactgataatgaa
tgagggattgaccccatcttttcaagct ttgcttcagcctaacttgactctcgtag
tgtttcagccgtttccatattaggcttg tcttccaccgtgtcgtgtcgtcaatctt
atttctcaggtcatctgtgggcagttta gtgcgaatggactcagaggtaactggta
gctgtccaagagctccctgctctaactg tatagaagatcaccacccaagtctggaa
tcttcttacactggcccacagacttgca tcactgcatacttagcttcagggcccag
ctcccaggttaagtgctgtcatacctgt agcttgcttggctctgcagatagggttg
ctagattaggcaaatagagggtgcccag tcaaatttgcatttcagataaacaacga
atatatttttagttagatatgtttcagg cactgcatgggacatacttttggtaggc
agcctactctggaagaacctcttggttg tttgctgacagactgcttttgagtccct
tgcatcttctgggtggtttcaagttagg gagacctcagccataggttgttctgtca
ccaagaagcttctgcaagcacgtgcagg ccttgaggtcttccgacttgtggcccgg
ggactctgctttttctctgtcctttttt ctccttagtgggccatgtcctgtggtgt
tgtcttagccagttgtttaagggagtgt tgcagctttatgattaagagcatggtct
ttccttgcaaactgcttggtttagaagc ctggctccaccacttagcggctctgtga
cctcggacacatttcttagcctttctgg gcctcgctcttcttcctcataaagtgaa
aatgaaagtagacaaagccttctctgtc tggctactgagaggatggagtgatttca
tacacataaagcacttaaaataatgtct ggcatatgatacatgctcaataaatgtc
acttacatttgctattattattactctg ccatgatcttgtgtagcttaagaacaga
ggtctttacaggaattcaggctgttctt gaatctggcttgctcagcttaatatggt
aattgctttgccacagactggtcttcct ctccttcacccaaagccttagggggtga
acgatcccagtttcaacctattctgttg gcaggctaacatggagatggcaccatct
tagctctgctgcaggtggggagccagat tcacccagctttgctcccagatacagct
ccccaagcatttatatgctgaaactcca tcccaagagcagtctacatggtacactc
ccccatccatctctccaaatttggctgc ttctacttaggctctctgtgcagcaatt
cacctgaaatatctcttccacgatacag tcaagggcagtgacctacctgttccacc
ttcccttcctcagccatttttcttcttt gtacataatcaagatcaggaactctcat
aagctgtggtcctcattttgtcaatcta atttcacagcctcttggcacatgaagct
gtcctctctctcctttctgcctactgcc catgagcagttgtgacactgccacattt
ctcctttaacgacccagcctgctgaata gctgcatttggaatgttttcaatttttg
ttaatttatttatttcatcttttttttt ttttttttttttttttttttttagggcc
gcacccatgggatatggaggttcccagg ctagggatccaatgggagctgtagctgc
tggcctacaccacagccacagcaatgca caattcgagccaatctttgacctacacc
agagctcacggcaacactggattcttaa cccactgattgaggccagggatcaaact
ctcgtcctcatagatacgagtcagattc gttaacctctgagccatgatagttgtta
gttactcattgatgagaaaggaagtgtc acaaaatatcctccataagtcgaagttt
gaatatgttttctgccttgttactagaa aagagcattaaaaattcttgattggaat
gaagcttggaaaaaatcagcatagttta ctgatatataagtgaaaatagaccttgt
tagtttaaaccatctgatatttctggtg gaagacatatttgtctgtaaaaaaaaaa
aatcttgaacctgtttaaaaaaaaaact tgactggaaacactaccaaaatatggga
gttcctactgggacacagcagaaatgaa tctaactagtatccatgaggacacaggt
ttgatgcctggcctcgctaagtgggtta aggatatggtgttgctgcagctccaatt
caacccctatcctgggaacccccatatg ccaccctaaaaagcaaaaagaaaggtgc
tgccctaaaaagcaaaaagaaagaaaga aagacagccagacagactaccaaatatg
gagaggaaatggaacttttaggccctat ctccaactatcacatccctatcaccgtc
tggtaagaaatggaaaaaatattactaa gcctcctttgttgctacaattaatctga
ttctcattctgaagcagtgttgccagag ttaacaaataaaaatgcaaagctgggta
gttaaatttgaattacagataaacaaat tttcagtatatgttcaatatcgtgtaag
acgttttaaaataattttttatttatct gaaatttatatttttcctgtattttatc
tggcaaccatgatcagaaatctttaaac aatcaggaagtcttttttcttagacaaa
tgaaaatttgagttgatcttaggtttag tacactatactaggggccaagggttata
gtgtgactattaaatcacagataatctt tattactacattatttccttatactggc
cccacttggatcttacccagcttagctt ttgtatgagagtcatccttaaagatgac
tttattctttaaaaaaaaaaacaaattt taagggctgcacccatagcatatagaag
ttcctaggctagcggtcaaattagagct gcagctgccagcctatgccacagccaca
gcaatgccagatctgagctgcatctgtg acctacactgcagcttgcagcaatgctg
gatccttaacccattgaacaatgccagg gattgaacacacatcctcatggatactg
ctcaggttcctaacctgctgagccacag ttggaactccaaagcagactttattctg
atggctctgctgatctctaacacgttat tttgtgccatggtgtttatcttcacttt
actcaagtcagggaaacacgaagagtct catacaggataaacccaaggagaaatgt
gcaaagtcacatacaaatcaaactgaca aaaatcaaatacaaggaaaaaatatctt
cactttcaaaatcacctactgatgatga gtttatatttccttggatatttgaatat
tagctatttttttcctttcatgagtttt gtgttcaaccaactacagtcgtttactt
tgatcacagaataatgcatttaagcctt aaatagattaatatttattttcaccatt
tcataaacctaagtacaatttccatcca g GTCTGTTAAATGCACAAAACACAACTGG Exon 6
Seq ID No 13 AAGTTAGATGTAAGCAGCATGAAGTATA
TCAATCCTCCTGGAAGCTTCTGTCAAGA CGAACTGG gtaaataccatcaatactgatcaatgtt
Intron 6 Seq ID No 33 ttctgctgttactgtcattggggtccct
cttgtcaacttgtttccaatctcatta- g aagccttggatgcattctgattttaaac
tgaggtattttaaaagtaaccatcactg aaaattctaggcaagttttctctaaaaa
atcccttcattcattcatttgttcagta agtatttgatgagacctaccatgtgtaa
acattgcactaggtattaagaaatacaa agatggataagatagagtcggcgtaaat
gagatgatataatgagacgttataatga aactcacaattccagttgggaaataaag
tccttcaaattccatgactcuctggcac acgttagaggctacagcttctgtgtgat
tctcatgctggctccacttccacttttt ccttcttcctactcaagaaagcctatag
aaatatgagtaagaagggcttaatcata ggaataaatttgtctctgttctaagtga
ttaaaaatgtctttatcagtataaaaag ttacttgggaagattcttaaaactgctt
ttacacactgttctagaatgactgttat ataaataaaaaagtagatttgatctaac
acaattaaatgacctttggaaatattga ctaattctcaccttgcccctcaaaggga
tgcctgaaccatttccttcttttgccag aaagcccccaccctttgtctgttgacct
agcctaggaaatcttcagatcacgttgt tagcacgaactggttacatgtgctgtac
aaatactatttaattcatctgattaaaa aaaaagagataagaagcaaaagtttgac
tatcttaaactgtttgcgtaggtgagag gacaattgaccatctactttatgagtat
gtaacccagaaacttaaagctccttaag ggagctaagtcttttggataagacctat
agtgagaccttttagcaaaatggttaag actgaatggagctcactagcgtgggttc
atatcctgatgctcaaacacgcaattaa atgactttaggtgggttagtctctgttc
cttagtttcctcaatgggagataatatt ggtagtagcgattttactgggttgttga
aagaacatctgttaaatgttcagaacgt gttacgacagagtacagagtaatgattt
gcttgtatatgtatgactcaaatagtct gccatatgccttgtgactgggtcctgtg
gagcaggaaggagggatttcccacccag cagaaagttgggtaaactggaaaataga
ctgaggccaggaaatgatgcaaagcgtt gatgttcactgccacggcaggtgaaggg
cagggccagagttgtcagtagggtcagg ggaggactggaaataaccaagacccact
gcacttttcagcctttgctccagtaagg taatgttgtgagagtagaaaattttgtt
aacagaacccacttttcagtacagtgct accaatactgtagtgatttcataccaca
tcccaagaaagaaaaagatggctcaatc ccatgtgagctgagattatttggtttta
ttgttaaataaatagcattgtgtggtca tcattaaaaaaggtagatgttaggaaag
tagaaggaagaagactctcacctacatt ttcatcactgttttggtatctgccagtt
gtcaccttggtccccttccccgcctctc ccctgcctcctcttcctccttctccttt
ttttggaatacaattcaggtaccataaa atttacccttttagagtgtttgactcaa
tggtttttagtattttcacatgttgtgc tattactatcactatataattccaggtc
attcacatcaccccccaaagaaaccttc taactattagcagtccattcccttcttc
cctcagcccctggcaaccactaatctac ttactgtctccatggatgttcctatatt
gaatcaagctagcataaaccccacttgc tcatggtcataattcttttttatagtgc
taaattacatttgctaatattcaattaa ggatttctatgtccatattcataaggaa
tattggtgtgtagttttctctttgtgtg atatctttgtctggttgggggatcagag
taataattactgctctcatagaatgaat tgagaagtgttccctccttttctattta
ttggaagagtttgtgaagtatattggta ttgattcttctttaaacatttggtcaga
ttcaccagtgaagccatctgggccatgg ctaatctttgtgaaaagttttttgatta
ctaattaaatctctttaatttgttatgg gtctgctcctcagacgttctagttcttc
ttgagtcagttttgttcatttgtttctt cctaggactttctccctttcatttggat
tatttagattgatagtaatatccccctt ttaattcctggctgtagtaatttgggtc
ttttctcttttttcttggtcagtttagc taaaggtttgtaattgtattaatctttt
caaataactaacttttttgttttgtttg ttttttgttttttgttttttgttttttg
tttttttttgctttttaaggctgcacct gaggcatatggaagttctcaggctagag
gtctaatcggagctacagctgctggcct ataccacaaccatagcaatgccagattc
aagctgcatctgcgacctacaccacaac tcggccagggatcacacccgcaacctca
tggttcctagtcggatttgttaaccact gtgccacgacgggaactcccgcccattt
tttttaacacctcatactttaacataaa gatgggcttcacatggactgatagctca
aatgaggaaggtaagactatgaaagtaa tggaagaaatgtagactatttttgtgac
ctagagattactgatacttcttgacttt tcaaacaatacttcaaaagtacagccca
aagggaaaaaagaaagaaaaaagaaaca cacatatacacaaacctagtgaataaga
tatcatcgatacactacagatttctatg aactggaagaccccatggacaaagttaa
agaacatatgatagtttgagtgattatt ttgcaatatttacaaccaatgagggaat
attatccagcttataggaggaagtaatg caaatcgacaagaaaaagataggaaacc
caatataaaaattaagaaaatacaaaaa ttaagaaaggatatgaactagcatttta
caaaagaaaaatctccaaaagtcaatca gcacatgaaaatatgctcaaaccaatta
ttagaaaactacagactgaagcaatgag gtgctttactttacatctttttgactga
taaaaagttagaaacaaaggtgatatca aatgtcagggataaaaggatatagaaat
cgtcatgcctgtggtgggagtatggccg gtgcagtcatgtgggaaggtaatctgac
agtggttaggcagagcaggtttatgaat acactgtggcccatcaatcccacgcctg
tttatgtaccaaagaaatcctgttgtgg cagaatctatgggtccacccctgggagc
atgaattaataaaatgtggcaccagggt gtgtgaaactccagctagagatgagatg
tccacatggcaacatgaatgcatcttag aaacatagatttgagtgaaaaagagtaa
gaaacagccgggaaacccaataccattt ataaaaattaaagatgcacacatacaat
gtagtaaatattttgcatgaactttcaa atggttgcctacagggggggagagtaaa
gaagagtagaaaacaaagataaagggag taagtaagtagctctgcctggactgaat
ataatgtgtcatgaactgagaaatatgg ttaacataatcctcttaacttgaggtcc
taaatgaatgaatgagtccactattcat ttacccattctttaatgtgtattgcatt
ataatccatttttttagaaccaacgaat tttgttcccataactactaatcagcctg
ccttttctccctcattcccttatcagct caggggcattcctagtttttcaaacgtt
cctcatttgaaccaaaaatagcatcatt gtttaaattatacttgttttcaaatacg
atgcttatatattccaagtgtgtttgcc cattttcttaggtggtagaaatttttca
ttctacttttctatctactcagattttc ccgttggaattatttccattgctattaa
acttagaagtcccccctgtgatatgcca tttttttcatactttttaagcacttggt
tgcttttctttgtgtctttaagcaccta gaatacttataaccattgcacagcactg
tgtatcaggcagcccttcctcttccact aatttatggtccttctcttagactatat
taaactgttatttaattaggatcctctc ttcgtccttatgatttaattattatagt
tttctaatatgtttttattataattcct cttcattattcctccctattaaaaattt
taatgaattccatttgtttgttcttcta gttaaatattaagtcataatccaaataa
cttagatgtcattagtttatgtggtcaa agtaaggataccacatctttatagatgc
aggcagttggcagatgtcatgattttct tcagtgcataaatgcaatttatctttga
gcaaggggcataaaaacttttatggtat tggctttgaaataatagttaagaactgc
agactcagtttttcctgcttttcttgaa aaagaacacttctaaagaaggaaaatcc
ttaagcatggatatcgatgtaattttct gaaagtctcctgtaattccttgggattt
ttgttgttgtttggtcggtttttttggg tttttgtttgtttgttttgttttgtttt
gttttgcttttagggctgcacctgtggc atatggaagttcccaggctaggggtcca
actggagctacagctgccagcctactcc acagccacagcaacatgggatcctagct
gcatctgtgacctaaccacagctcttgg taatgccagattgttaacccactgagca
atgccagagatcgaatctgcctcctcat ggacactagtcagattagtttctgctga
gccacaatgggaattcccaattccttgt atttttgaactggttatgtgctagcata
taattttgtttcttgaatctttgtgggt ttttttttttttttttttttgtctcttg
tctttttaaggctgcacccacagcatat ggaggttcccaggctagaggtcaaattg
gagctacagctgccagcctacacaacaa ctgcagcaaagtggggcccaacttatat
gacagttcgtggcaatgccggattccta acccactgagcagggccagggatcgaac
ctgagtttccagtcagtttcgttaacca ctgagccatgatagtaactcctgtttgt
tcagtcttgaacctcctttttaattctt tattccttgagggtgaaataattgccat
aataatactatcatttattacatgcctt ctctgtgctaggcatagtgacactttag
gatttattatatcacttaatccctacaa caactctgcaaagtatgtatcataatcc
tatttgacagatcaggaaattgcagccc aggatgcagataatatgcatccatcaca
agtgactagatatagtccctctgctatt cagcagggtctcattgcctttccattcc
aaatgcaatagtttgcatctattgtata tgtgttttggggtttttttgtctttttt
tttttttttgtcttttctggggcctcac ccttggcataggtaggttcccaggctag
gggtcaaattgaagctgcagctgccagc ctacaccacagccacagcaactcgggat
ctgagcctcatctgcaacctacaccaaa gctcacggcaacaccggatccttaaccc
actgagtgaggccagagatcaaaccggc aacctcatggttcctagtcggattcatt
aaccactgagccacgatgggaactccct aaatgcaatagtttgctctattaacccc
aaactcccagtccatcccactccctcct cctccctcttggcaaccacaagtctgtt
ctccatgtccatgattttcttttctggg gaaagtttcatttgtgccatttttcatt
ttacgggtaatttttacttcagtttctt ccactagcagttgtcttaaagtgagtat
aattaatattcatttggaaaatgtaagc aaaacattttttaaagggccatgcccac
agcatatgaaagtttctgggccaggggt tgaatccaggctccaagttgcagctgtg
ccctacactgcagctgggcaatgctgga tcctttaacccactgtgcccggctaggg
atcaaacctgcatttccacagctacccg agccattgcagttggattcttaacccac
tgcactacagtgggaactcccacaaaac attttttaatgtcctttgaataaagtag
gaaagtgctcgtctttgagggcagggcg gcaatgccatttccacaaggtttgcttt
ggcttgggacctcatctgctgtcattta gtaatgaataaaattgctgacagtaata
ggattaactgtgtgtggagatagccagg gttagagataaaaacactggagaagtca
aataagttgctcgaggtcctctagctaa taagctattaagtgggagagtgagggct
agaaacaggccatctgtctcccaagcac atgtccattagtggtttgctgatagcct
tccagaacaacagagaggactctcaaac atggtcttgcctccctccaattgatccc
ctccatgtgcctcacagcgggtctttct aaaattaagttctgattttaattctccc
ttgctatagcacttaggtatggctttca gccgtgcaataaaaagcaggcaagagtg
gctcaatcatataggaggttgtttttct tagatcccaagcaggtaatcctgggcat
tatggttgttctgcgtttatcaaggagc caaattctctatcacctcctgttctatc
ctcctcagtatctggctctattcttcag catctcaagatggcttgtgctcctccaa
gcatggcagtcaaattccacacaagagg gggaaatatgaagggcagacagtgctgg
tctcctgagctgtccctctttgtcgggg aaatgtattccttcaagtcccgtgagac
ttctgaagtagacgtctgcttacgtctc acccaccagaactatgtaaactgcacat
agtgctaggtctacatagccactcataa ctgccagggggtgggaaatctttaaata
ggtgtaccaccacacaattaggatgcta atagtaagggagaaggagagaataggtt
ttgcgcaagccaccagcatgcctgccac aattgcttaaaattcttcattgacccct
cattgccacaggatgaaatccaaacgcc
ttcttagttgggaatctgacctacctgt ctctcccacctggttcagacaccattct
ccttggtcataaaattccagtcatttgt gaacatccagctcccccatgcctccatg
cctttgcacatgctgttcttttatcttt tatgttgtccttttatcttttatccaaa
agagatatcccatcatcacatctctttt gtcagcccccaaatactttgtctttcaa
gttcagctggaggattacctcctatttg aaatcagctttgtctcttacaaccaaac
aaggttttccttccgagacactcccaca gcaccttgaactcatctctatcaatcat
tcatttgattgtaatgaagttgttggtg gtatgcctgtgtctctgacacatctgcg
atctcatgagttccttaagtggaatgtg aatagcgggatgaacagtattggtcttc
agccctcatctctgcagatgttgcttga cccaaatgagcgttgccttttattttga
ttttgctttgatttgtctactccatgta cttgagccatgcatttctgtcttagcga
tgctttttaaaagtcattttttggttga ttatccagatttgtccacctttgcttct ag
TTGTAGAAAAGGATGAAGAAAATGGAGT Exon 7 Seq ID No 14
TTTGCTTCTAGAACTAAATCCTCCTAAC CCGTGGGATTCAGAACCCAGATCTCCTG
AAGATTTGGCATTTGGGGAAGTGCAG gtaaggaaatgttaaattgcaatattct Intron 7
Seq ID No 34 taaaaacacaaataaagctaacatatca
atttatatatatatatatatatatattt ttttttttttttacatcttatattacct
tgagtattcttggaagtggctagttagg acatataataaagttattctgaagtctt
tttttttctttttccatggtgagcagtg gcttgatgtggatctcagctcccagacg
aggcactgaacctgagccgcagtggtga aagcaccaagttctagccactagaccac
cagggaactccctattctaaattcttga gcacattatttaggaacctcaggaactt
ggcaggattacaggaaatatatctagat ttaaaaaaaaatcttttaacagaggtcc
caaaggagagtcatgcacagctatggga ggaagttcagaaactgcccttgctacca
gatcactgtcagataaaatggccagcta catgtttctgcacattgccctaagatct
ttacaaacttttctgtgcatttttccac ttttaaaagaaaatttcggggttcctgt
tgttgctcagtggttaacgaacccaact agtatccatggggacaggggttcgagcc
ctggcctcactcagtgggttaagaatct ggcattgctgtggctgtggcgtaggctg
gcggctacagctcagattggacccctag cctgagaacctccatatgccgcaggtat
ggccctaaaaaaaaaaaaaaagagagag agagaatttcctccagaaaaaacacttt
ggtagtttgggagaagtaaacaaccaaa aattaatttttctggagtattcgggaag
cttgtaaaaatgggctcttacttttttg aggagacaaatgggaacctacccagaag
aggcacaatcacctgcatttgatttctt gacctctccctaccttctttgctggctt
tccacatttggatttctgtgaccttatc tctgctccttggtgttttcatttttcct
gtggacgtgccagactatgggaagggag taaggcgttgatttagaatcctgtagtc
tctgcctgtctctagtcattgttttcac ccttctcaaaggaccttgacatcctgag
tgagtccgcaagtaatttaggggagaag ccttagaagccagtgcagccaggctaca
tgactgtgtccacccactggaaccagtc atttttatacctattcacagccccccta
ccatttaaatccccagaggtctgccata acatctgtaactccctttcctggtaaat
tgtgttctaaaagactggtaacaaaaga tattctgtggtacagagcataattaaat
acctgggagctgatttgagtggggtaaa tcaactggtttgacccctaaaacccacc
atgagcatttctgttctaataaagtaat gcccgtgctgggaattgtgttctacgga
aatgctcctgctgtgtctttcttgagtc ctgtgtcattgaacatgcttaggagcaa
aggtcccccatgtggcttgtctgctaac cagcccagttccttgttctggctggtaa
tgatccgatcatctgaatctcactgtct tccaacag ATCACGTACCTTACTCACGCCTGCATGG
Exon 8 Seq ID No 15 ACCTCAAGCTGGGGGACAAGAGAATGGT
GTTCGACCCTTGGTTAATCGGTCCTGCT TTTGCGCGAGGATGGTGGTTACTACACG
AGCCTCCATCTGATTGGCTGGAGAGGCT GAGCCGCGCAGACTTAATTTACATCAGT
CACATGCACTCAGACCACCTGAG gtaaggaagggtgagccctcaactccga Intron 8 Seq
ID No 35 agaaaatgctgcaataaaagcactgttg gttttcagctttttttgtaatcactgct
cattctgaggtagattcgcttgggctga taaaaagagaactaattcagataatgct
tgcatttgcatagcctctttttttaaaa actttttttttttttttttttggctttt
cagggctgaacctgtggcatatggaggt tcccaggctaggggtcgaatcagagctg
tagccccgggcctatgccactgccatag caacatgcatagcctcctttttaaagtg
ccttcctgttttataccattgggatgtg agaagagctattgtggaaangagcatgg
ggtnataaccctggacctctcacgtcct accctcaggntagtgggaaaactctgag
tttaaggacatcaaagtgactccttttt agttacattatggnggaatcagcncata
tttttacaaggggcggagngtaanctgt tggagtttacaagacatatggtggcatt
gcaactacttaaccctactattatagca caaaagcagccatagtcggtcctgaagg
agcctgatgccttcagctttataggcaa tgacgtgtgaatatcacaaacagtttcc
tgtgtcaccaaacatgattgccttttga tttccctttcaaccctttaaaaaaaggt
aaaagcccttcttagcattcagcagcag gtcgctgtgttttgccaactcctgatct
gtagcatttcgacaacactgagctctca acttttgaaccctgagtccaccacatcc
ttcagtgaaaccagagccatgtgatact aaggatagaaacggaaacttcctgaatc
caggcgatcaaataggagggagaaagag gaactttcattgacaaaaccacaaatat
tgtgaatggactgttacaaatattgtga atgctcctattcccaaccccctggcttc
attacagggtcctatgtgttcatcctta ttgagaaatttgtattgctactgccagg
ttgccaatacccagcggtgcccatggtg ttctaaaatgaagcaatttcaactttat
ttttttttcctgtgactttacatgacaa gttcacatgaaggatatactttgatagt
aatgtccatggttagggaatatacattg tttgctggttgactggcccctggatttt
tctattgaaagtccatgagatctcgaag gcacaggtgtgttctctcgctttttaag
gaaagggtttaaaaacttaagtaattaa cagctttagtaacaaattacctataaca
cacttaaaaaccgaataccacccactgg agtattgtgctacgattaaaaatctact
tgtctactacatgatatctttgtcccac agaaggttctggaaccaaacttgtaatt
tcaggattatgagagccctgagttcacg cattgtgtaataactatgttgtgtggta
gtcaatttgtacagcttgcttagagaga acaatgtcaagttaaggaggcgattgct
ttatagtgcctgtcacaagatgccattg ccattgtcctagcaagagatattctatg
ggagtatactacattttagtgaggataa gaactttttatggcatttagtccggtca
tttcccaaccactgtcctgaaaaccaat ttcattttgatttcaggggcttgtgtgg
gcaaagttgccaggcattaaaaagccac ttctcaactgtagtatcacaatgcttta
gttgggtagtgtattgcagatagcttat ggctgaaaagttaccaagccttgcagtt
ttcactcctttgagtttatttccttgac agaattgaccctgagttttttgactctt
acctgctcaactaataaacaccagagtc atttatctccattgctcttgtctgacct
ttatttaccgaataatgccttatgggtt cacaaaaacaaggggggagggggccagc
atgccttagaaactgtctttagtcaaga aatgngattttattatgtaaatatatga
gtattataatagatagtgttattaatag acaccagcaagaattgtcaataatttaa
aaatcacaaattaaaatacatccatgtt agnatcatttatcctaactcccaaagcc
ctttaaagtggaagatttagatgttaac ccagagattaaagacatgttcaaagaat
ccttgatttttttttgaatcccttgttt ttagagaagaaaacctaatgattttccc
cctctggattctacatattaaatatagt tttggaacttgaatattagtatggttaa
taagtgctgatatgctgattttgtttat atttttcttatgagtaaatatcctatat
caccagacattatagtctatgtacaaat atgattcttaaacctgatagcacattca
ttagagttggaattgccttttttttttt tttttttacagttgcacctgcaacatat
gaaagttcccaggctaggggttgaatcc aagctgcagctgccaccctacattacag
ccgtagtaacagcagatccgagctgcat ctgcaacctatgctgcagctcagggcaa
tgccagatccactgagtgaagccaggga tggaacttgcatcctcatagagacaacg
tcgtgtccttaacccactgagccagaac aggaactccagaatttcctttcaataga
agaagcaccaagtttaggatcagaaagc ctgaatttgaataccaatttactatttg
ttagtcatatatttctgagtgtgtttcc tcatttattaaaagcagactaaaagatg
agagggtcttttgttgagaatcaaatac aataacatgtgaaagtgtgtaacactat
gattgaaatatacctacacagccattta tttgtttattgttcatgttttgccaccc
acacagtagtatataatccttttatgta ataaatgctaataatgaaagttggcaac
ttatgtaagtactcaaaatgctggaggt catgggatactgactgggatactacaga
ggtaatgtcatttcctctgcgctaaact tattgtcttgtagttagggactgactct
ctttaggacaaggagttcattctgtata ccatggctatcacccttcgaagttgaaa
aactgccccagggtgggcacccatccgt tctcttagatatatggccgagacctttc
tctcactgggagggaaccacactgagga atgagaaaaaaaaaaggaaaatcaagat
gaaaccagaaacctctttggcataactt ctccactctgtactttttgttagaacta
cccttgcacaaagcagcatcagtgtgga agacagaatttgcacacctggtttgata
tacatgccgtggtatatgggatgttcta acaataaagaggactctcccaggaaatc
tcctcactgttatagtcagccttgagga aagagctcttcttttggactctggggag
agtctagtttttcagttccttgcttctc ggtcaacgtgttggtgtaaggatcacac
tctctcttatactagataattctatttt ttcacctttcaacctgtctatccttctg accctag
TTACCCAACACTGAAGAAGCTTGCTGAG Exon 9 Seq ID No 16
AGAAGACCAGATGTTCCCATTTATGTTG GCAACCGGAAAGACCTGTATTTTGGAAT
CTGAATCAGAGTGGCGTCCAGTTGACTA ATATCAATGTAGTGCCATTTGGAATATG GCAGCAG
gtctgtgttctttccacatgtttgggtt Intron 9 Seq ID No 36
atcctttctgggataaatttgaggcgag atagaaactttaagactaaagaaa- caat
ggcctactttttttgtacatggtcctgt gtaaatctctatttgagctgaaataa- ga
tggtcttcctctccaattatccatggta tgactctgatggataacaaatccagttc
tgaaaaaaggggatttctttccagaaga gaggacagtttcttcaaatattgaatta
aaagcaaaatagatgtaaaccgttgttg gttttattgttgaattccag
GTAGACAAAAATCTTCGATTCATGATCT Exon 10 Seq ID No 17
TGATGGATGGCGTTCATCCTGAGATGGA CACTTGCATTATTGTGGAATACAAAG
gtattttcttgccctcatcagcatgaaa Intron Seq ID No 37
ttgctcttggtagaaaggataataatag 10 ttatccaaaacatcatcctatgttcatc
tgtttcttccctcttcattttccataga gtacagtatattctatctctgtcttagg
aaaatggactgtcattcatataatctta cagagaatcaattagtaatgtactctat
gccgtgacaggtgcgaaggttttttttg aaggcaacagataaaaatatcctatatt
tcacctattgtaatttccttaaaactga cattattgaataaatgttttactttcat
cttgaatattattatgttatggaatcat acactttaccccaataatcatcgaaaag
aatttccaaaaggttgagagagttgtgt tgatctgattactttcctctgcatcctt
tgagcttaacctttgaatatagtttgct aaggaaagtagtctgtttatgatcctgg
agtggaatcaggctaagtgtcctcattc agaacccactgaatcagacagaatgaat
ttatttccttgaaagttcaaaatgtgtc actcaagagtataaattttcaaatctta
ctctctcttttccttggatgtgagcaat tcttcgataattgaatgaggcagattat
atagacttacatggaagactgttggcct gagaattcaaactatggtgttcaagact
tcacngngagtccgatgccatttgtttc ccacag GTCATAAAATACTCAATACAGTGGATTG
Exon 11 Seq ID No 18 CACCAGACCCAATGGAGGAAGGCTGCCT
ATGAAGGTTGCATTAATGATGAGTGATT TTGCTGGAGGAGCTTCAGGCTTTCCAAT
GACTTTCAGTGGTGGAAAATTTACTG GTAATTCTTTATATCAAAATGATGCCAA Exon 11a
Seq ID No 19 GGAGTTGGCATGGCACTTTGCTAAATGC
TGTGTGAATCAATACAAAGATAATTAGG ACATGGTTCTTCCTCACAAGAGGTGTGC
AATCTTATTGGGAAATCATACTTGCAAG TCACAAATATAGACTAAAGTTTCCAGCT
GAGAATATGCTGATGGAGCATGAAACAC TAAGGAGACAGGGAGAATCTCAGGAAAA
ATCAAGAATAATTTGGATCAAATGGATT CCTGACATAGAACATAGAGCTGATCAGA
AAGAGTCTGACATTGGTAATCCAGGCTT AAGTGCTCTTTGTATGTGGTTCAGAACA
GAGTGTGGGCAGCCTGAGGGGGATACAT ACCCTTGACCTCGTGGAAAGCTCATACG
GGGGAGGGATGAGGCTAAGGAAGCCCCT CTAAAGTGTGGGATTACGAGAGGTTGGG
GGGGTGGTAGGGAAAATAGTGGTCAAAG AGTATAAACTTCCAGTTACAAGATGAAT
AAATTCTAGGGGTATAATAACAGCATGG CACTATAGATAGCATATTGTACTATATA
CTGGAAGTGCTGAGAGTAGATCTTACAT GTTCTAACCACACACACACACACACACA
CACACACACCACACACACACACCACACA CACACACGTGCACACAAACAGAAATGGT
AATTATGTGAGGTGATGGCGGTGTTAAC TAACTTTATTGTGGTCATCATTTAGCCA
TACATGCATGTCATGAAATCACCATGTT GTACACCTTAAAGTTATGTAATACTAGA
TGTCAGTTATATCTCAAAGCTAGAAAAA ATGTGGGGACCAAGGCAGAAGCTCTTCT
GCTCTGTGTCTAAGGGTGGTTCTGGGGC TGGGATGGGGAGGATGGTTAAGTGGTAT
ATTTTTTTCATACCTTTGCTCAGTACTA TCATTGTAAGTGTTCAATATATGTCTGC
TTAATAAATTAATGTTTTTAGTAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAA
gtaattctttatatcaaaatgatgccaa Intron Seq ID No 38
ggagttggcatggcactttgctaaatgc 11 tgtgtgaatcaatacaaagataattagg
acatggttcttcctcacaagaggtgtgc aatcttattgggaaatcatacttgcaag
tcacaaatatagactaaagtttccagct gagaatatgctgatggagcatgaaacac
taaggagacagggagaatctcaggaaaa atcaagaataatttggatcaaatggatt
cctgacatagaacatagagctgatcaga aagagtctgacattggtaatccaggctt
aagtgctctttgtatgtggttcagaaca gagtgtgggcagcctgagggggatacat
acccttgacctcgtggaaagctcatacg ggggagggatgaggctaaggaagcccct
ctaaagtgtgggattacgagaggttggg ggggtggtagggaaaatagtggtcaaag
agtataaacttccagttacaagatgaat aaattctaggggtataataacagcatgg
cactatagatagcatattgtactatata ctggaagtgctgagagtagatcttacat
gttctaaccacacacacacacacacaca cacacacaccacacacacacaccacaca
cacacacgtgcacacaaacagaaatggt aattatgtgaggtgatggcggtgttaac
taactttattgtggtcatcatttagcca tacatgcatgtcatgaaatcaccatgtt
gtacaccttaaagttatgtaatactaga tgtcagttatatctcaaagctagaaaaa
atgtggggaccaaggcagaagctcttct gctctgtgtctaagggtggttctggggc
tgggatggggaggatggttaagtggtat atttttttcatacctttgctcagtacta
tcattgtaagtgttcaatatatgtctgc ttaataaattaatgtttttagtaagtaa
tctctgtttagtaatgtgtcagaaatgc cctacttgcaataggaagaaaacctgtc
cagtcccttccttttttctgtaagtctg atttcattgcctcccagaatgcatcacc
atgtgagagatagagggaaggtgctgtc cttatggggttaacagtgtgactaggga
ggcaaaatatacctactaaagggtggta gcataattcagttcttatgtgagtatgt
gtatgtgtgtgagtatgtgcacatgcac atacattttaaaaggtctgtaatatact
aacatgttcatagtggttacacctagct tataggtaacattttttcccctgtatcc
ttgtttgtgtttatcaaattttcataac agtaatggtagaaggagtacctgacatg
gtaccatacatgctnggncctgcctaat ttctcnatttcctttattgcccataccc
ccattgcttgacaagcataagtccatac tggcttgttttcgttcctcagactcagt
acaccatgtagctccatgccctgggtct tgtatgtgctatttctactgcttagagt
gctattgcccctgaccaccacgtggtca gcaacttctcttctgcgtctgtgtctat
ggtctatgattccagatgtcatcttcac taactacccttctaatatgcccttccat
cccacccgtcctcatccttaccccagcc actctctatttggtggctctgattttct
tcctagctcatcactctttgaaatgaac ttatttacttattcaatatttgcttctt
tcactagaatgaatgctccatgagagca gggacctgctttatcttgctcgccactg
tattcacagtgcctagaactacgtctgg cacatagtaggtgctcaataaatatcga
tcaaatgaaagaatgagcaaacgaacaa atgaacaacacgtgaggtaggcatcatg
attccatcaacagaggagaaaaccagac ttaaagnaatgaagtggnggagctgcat
ttgatcttgactgactccaacatccatg ctcttgaccactgtgcatctccagagtg
taatgaacatactttacttttatattcc accaaaataacaaagccatgcccatgtt
agtagagagttaatcgacagtgccctta aaatatgcatgcacccagggtacaacta
tgcatgctgccctgtgttttcagttgga tccaaatgaattgccgtaaacaaagagg
ggattcaatgtctttgactagtttggga tattttcctagtaaccaactttgcaaaa
taaagccactaatgacaaggagctttgt tctacttctgcatcactcaactgtcaat
ttttatctcttgcaagacttctaatcta ctagaacttttgtttttctgtgatttct
gaacagagaagactaatccaaaccctgt cattccag AGGAATGGAAAGCCCAATTCATTAAAAC
Exon 12 Seq ID No 20 AGAAAGGAAGAAACTCCTGAACTACAAG
GCTCGGCTGGTGAAGGACCTACAACCCA GAATTTACTGCCCCTTTCCTGGGTATTT
CGTGGAATCCCACCCAGCAGACAA GTATGGCTGGATATTTTATATAACGTGT Exon 12a Seq
ID No.21 TTACGCATAAGTTAATATATGCTGAATG AGTGATTTAGCTGTGAAACAACATGAAA
TGAGAAAGAATGATTAGTAGGGGTCTGG AGCTTATTTTAACAAGCAGCCTGAAAAC
AGAGAGTATGAATAAAAAAAATTAAATA C gtatggctggatattttatataacgtgt Intron
Seq ID No 39 ttacgcataagttaatatatgctgaatg 12
agtgatttagctgtgaaacaacatgaaa tgagaaagaatgattagtaggggtctgg
agcttattttaacaagcagcctgaaaac agagagtatgaataaaaaaaattaaata
caagagtgtgctattaccaattatgtat aatagtcttgtacatctaacttcaattc
caatcactatatgcttatactaaaaaac gaagtatagagtcaaccttctttgacta
acagctcttccctagtcagggacattag ctcaagtatagtctttatttttcctggg
gtaagaaaagaaggattgggaagtagga atgcaaagaaataaaaaataattctgtc
attgttcaaataagaatgtcatctgaaa ataaactgccttacatgggaatgctctt atttgtcag
GTATATTAAGGAAACAAACATCAAAAAT Exon 13 Seq ID No 22
GACCCAAATGAACTCAACAATCTTATCA AGAAGAATTCTGAGGTGGTAACCTGGAC
CCCAAGACCTGGAGCCACTCTTGATCTG GGTAGGATGCTAAAGGACCCAACAGACA G
gtttgacttgaatatttacagggaacaa Intron Seq ID No 40
aaatgatttctgaattttttcatgttta 13 tgagaaaataaagggcatacctatggcc
tcttggcaggtccctgtttgtaggaata ttaagtttttcttgactagcatcctgag
cttgtcatgcattaagatctacacacca ccctttaaagtgggagtcttactgtata
aaataaactattaaataagtatctttca actctggggtggggggggagactgagtt
ttttcacagtcctatataataattttct tatcctataaaataattaggagttcccg
tagtggctcagcaatagcaaacccgact agtatcgatgaggatgcgggttcgattc
ctggcccccctcagtgggttaaggatct ggcattgccgtgagctgtggtgtaggtg
gcagacacggctcagatcccacgttact gtggctgtggcataggccagcagctcca
gctctgattagacccttagcctgggaac ttccatatgctgtgggtgtggccttgaa
aaaaaataaataaataagataattactc aaatgttttccttgtctcagaaccttac
ttcaggataaagagtgagaaagtttttt ttatgaagggccattattacagctcaaa
aataagttgtcttcagcaagtagaaagc aataagcctgagagttagtgttcctatc
agtgtaaatattacctcctcgccaatcc ccagacagtccatttgaacaattaacgg
tgccctgggagtacagttcagaaacatt aatgtggatgttccagacctgtattttt
ataagtaccttgagccggatggaaccat cattcctcaccattatttagaagtggac
tgtgactctgttggagatcagggcacac ggttaccaaaagcacacccttctcctgg
ccttacctttgcaaagctggggtctggg acacagtcagctgattatacccttttac
taacttcccacagctcaaatctggtcaa ttctccttcacaaatctcttaaaaatcc
atcactcacctccagcctcttctgctgt ggccttgattcagcctctcacaattttt
ttttaaccagaattctggcagtggcccc tgacttgcctctgtgctcccagccccgc
tgtcctctgatccatcctccatgccagc ctttttcaatctgctggtcacgattcat
tgatgggttaggaaatcaatggcatcac aactagcatttagaaaaaggaaataggc
gttcccgccgtggcacagcagaaataaa tccgactaggaaccataaggttgcgggt
tcaacccctggccttgttcagtgggtta aggatccggcattgccgtgggctgtttt
gtaagtcacagacatggctctgatccgg cattgctgtggctctggcgtaggcctgc
agcatcagctccaattagacccctatcc tgggagcctccatatgctgcaagtgcag
ccctaaaaaaaataaaaaaataaaaaaa aataaataaaagaagtagacaaattgta
tagaacaaccctgagtatgttgcctgag cacatataacaagggtaagtattatttc
aggaaactctggtttcacagatactctt ggcatatggacccctagagtcctgatgt
aaaatatattcttcctgggatcttaggc aagaagtttgaaagctccaactctgcac
tgctgccaaagaaatgatttttaagtgc aaaactcttcccgttcccttccctgtat
aaaattccataggatctctccagtgcct ctaggataaaggcagttttcattctcta
gttcaaggtgagagaagattttaattat ttcacgttttagtggggaattcaagagt
ctggcacctgacatttgctgaactctct ccattatccctctctagttccccagacg
catcctatggtagaaattcgcaaactag agtgagcgtcagagtaacccaaggaaac
tgggtaaatgcagctccctgggctctac cccctgagattctgattcagtagatctg
aagcagagccctggaatatgcatatgca tcattgtgtcacaccaagcattctgggt
aatgagagttgatgttaggttctcagta gtaagacaagtatagagattccggggga
ctgagtgctcagctctgccttggggagg agggagagggctaaagagaacaggagat
ggggacagggaatgctcaacctccaatc ttaggcatttgagctatgtcttaggggt
caggaggaggttaccaatatagtgatta agagattgaggttccagtcagagggata
tgctggagaaggggggtgaaaataatgt cataggtttggtgagtgcagatactttg
agttttttaatatttttattgaaatata gttgatttacaatgctcttagtgagtac
aattactttgaataagtgcatagatgta tgccattcttccagaaatgatttattga
gctcctttgggcatcatgctaagtacag gggaaacagctgtgaagaggtccttccc
ttatgaagtcattcatccccttcagtaa atgaaggtaaaggaaaaggatgagacag
ggacgccgtgttggaccagggtcagaaa ggccttataagaccttgcctggagggca
aggaacttgcctgtgagtaaggagagct tgagaaagcgataaagcaaagaaggaac
attactgcattgtgttttagaaaaacca tgtcctggggaagaactcctagagtcag
gggggccagttgggagactgtgcttttt tccaggaggagataagtgaggctgctgg
ctgagatggagcaaggatttagagaagc agatatgagattcatttagaagttagac
attttaggatctgacacataatttatca ccaaaaccagtgcatctctggctttggg
ccaccagttttggagaagtggaatgtag ggacctaccattacctgccaatctttac
tacacagatgcctatttccctcctcata ttcctttctccagatcacgtcctattct
attgccaggactcaagattccaccttgc atgcagtgatccatcttcacactggatg
gacagctctagggatgtcagagcacact cttgtccatactgctgactgggtctcct
gtcagcccatctgtctatcagctgtggt attattagtataataagagggctgtata
tgagagacacaaaattctaggtgtagct caaagataggctagagttattcctatgt
acaacaaatatttatgggaccccttctg tgtactgtcatggttgctgctttcatca
tacttgtagtctaatggaggtgggggca gggcaggaataagcggatgtccacaaaa
tcagtaagaccacttatattcaacattt tcataatttagttatttgagcccaaagg
gtccacatccgtggtattccaacttttt tttccccggacatggatctttatctttt
tttttttttcttttttgcggccagacct gcggcatatggaagttcccaggccaggg
gttgaatgggagttgcagctgcctggtc tacaccacagccacagcaaggtgggatc
tgagctgcatctgtgacatacaccgcag ctgaggtaacaccagattctgaacccac
tgaatgaggccagggatggaacccgtct ccttatgaacactatgtcatgttcttca
ccctctgagccacaacgggaactccaga cttcgtctttaaatgtattctgacttgg
agagctatcacactaagcaattaacagg agctgacctggtttaggctggggtgggg
ccctactcctcaatgttccctgaggcac atctgtgggacccctgggcatcatctat
ctgagcagccttagagctgctcatccag ttgactgttgatgtagaagtgcaaactt
ctgccttccttatttgttgctttctttt ttcattgttctctcccctttgtgtcttt aag
CAAGGGCATCGTAGAGCCTCCAGAAGGG Exon 14 Seq ID No 23
ACTAAGATTTACAAGGATTCCTGGGATT TTGGCCCATATTTGAATATCTTGAATGC
TGCTATAGGAGATGAAATATTTCGTCAC TCATCCTGGATAAAAGAATACTTCACTT
GGGCTGGATTTAAGGATTATAACCTGGT GGTCAGG gtatgctatgaagttattatttgttttt
Intron Seq ID No 41 gttttcttgtattacagagctatatgaa 14
aacctcttagtattccagttggtttctc aataagcattcattgagccttactgact
gtcagacggagggcgtattggactatgt gctgaaacaatcctttgttgaaaatgta
gggaatgttgaaaatgtagggaatgaaa tgtagatccagctctgtttctcttttgg
aggattctttttcctccatcaccgtgtc ttggttcttgtttgttttgggtttttgt
gggtgttgtattgtgttgtgttggttat ggcagtgacagctatttaaactgtgaaa
cgggggagttcccgtcgtggcgcagtgg ttaacgaatccgactgggaaccatgagg
ttgcgggttcggtccctgcccttgctca gtgggttaacgatccggcgttgccgtga
gctgtggtgtaggttgcagacacggctc ggatcccgcgttgctgtggctctagcgt
aggccagcggctacagctccgattggac ccctagcctgggaacctccatatgccgc
aggagcggcccaaagaaatagcaaaaag acaaaataaataaataaataaataagta
agtaaaataaactgtgaaacggggagtt cccttcatggctcagcagttaacaaacc
cagctaggatccatgaggatgtaggttc gatccctggccttgctcagtgggttaag
aatccagcgttgctgtgagctgtgatgt aggtcgcagatgcagcccagatcctgca
ttgctgtggctgtggcgtaggctggcag ctgaagctccgattcaacccctagcctg
ggaacatccatatgctgcaggtgtggcc ttaagaggcaaaaaaataaaaaaataaa
aaataaataaattgtgggacagacaggt ggctccactgcagagctggtgtcctgta
gcagcctggaagcaggtaaggtaaggac tgcagctgggtaaggactgaattgcacc
aactgggaagtaagcctagatctagaac ttaagttagccctgacatagacacacag
agctcaccagctaagtggttcagcttat aagctggtcactgaaactgaggatgtcc
acaaaagcaaaataagtagcaacaggca gcgggatgcaagagaaagaggaggccta
aaatggtctgggaatccctgccatacct atattttatcctacttatatttagtgcc
tgaatgtgtgcctggagagcaaagttta gggaaagcatcgggaaatgcacagtatt
catacccttaggaacaaagatcagttac ctccagggtaaagactatttccaagttt
aaatttcaacccctgaacattagtactg ggtaccaggcaacacttgccatcctcaa
aatcaatgaatcctaaaattcaacctgg gggtcagtgacagtctgtgacaaagttt
ttgctggtcagtaacgaaataagtatga gcaccatctgagtatggtcaccaagatg
tcaactctctttcctttggacgaacatt attccaagattaggtcctttctattttt
gaggtgtgaaaacatctttcctttcata aaataaaaggatagtaggtggaagaatt
ttttttgttttttggtctttttgctatt tctttgggccgcttctgcagcatatgga
ggttcccaggccaggggtcgaatcggag ctttagccaccggcccacgccagagcca
cagcaacacgggatccaagccgcatctg cagcctacaccacagctcacggcaatgc
cggatcgttaacccactgagcaagggca gggaccgaacccgcaacctcatggttcc
tagtcggattcgttaaccactgcgccac gacgggaactcctaatgatactctttta
tatttagctactatgtgatgatgagaaa cagtccacattttattattttttagcca
atttgatatctcattactaagataatga taattttctctataaattttatttaagt
tagtgttatgaagtggttttgctagtgt agaaggctaggatttgaattcagttcaa
gaaagaagagagggagggagggagaggg atgggtagagggatggggcagtgggaga
gagcaaagaggagagacagtttttgtat taattctgcttcattgctatcatttaag
ggcacttgggtcttgcacattctagaat tctaaggaccttgaccgccagattgata
tgcttcttccctttaccatgttgtcatt tgaacag ATGATTGAGACAGATGAGGACTTCAGCC
Exon 15 Seq ID No 24 CTTTGCCTGGAGGATATGACTATTTGGT
TGACTTTCTGGATTTATCCTTTCCAAAA GAAAGACCAAGCCGGGAACATCCATATG AGGAA
gtaagcaggaataccagtggaagtgcc Intron Seq ID No 42
cctttcttccttccttcctaaataaac 15 ttttttattttggaacaactttagagt
tacagaaaagttgcaaagatattatag acagtagtgtttatatatatatataaa
tttttttttgctttttatgaccacacc tgtggcatatggaggttcccagtctag
gggttgaattggagctacagctgccag tctgtgccataaccacagcaatgcagg
atctgggccacgtctgtgacctacacc aaagctcacagctggattcttaaccca
ctgagcaaggccagggattgaacctgc atcctcgtggttcctagttggattcgt
ttccgctttgccgcaatgggaactcca aattattgttaatatcttactttactg
gggtacatttgttacaaccaatactct gatactgaaacattactgttaactccg
tacttgcttctttttgagtcatttgca aagactggcttcttgacctgcttcctt
ccaaacagctggcctgcctatgctgtt ctcagacctgcaagcactgatctctgc
cccccttgccttctctccagtggtgtc tccttccccaaacaaacccagtgtggc
tctggaaagggagttaagtcaacataa accaacacatattttgttgagctccaa
ttttgagcaaatccctcacctacggca gacaggcatgatgttaagaactagggc
tttggacacaaggtcaagaccaagaag ggttcctcacccctactgattcagata
accaataatgaggctttgaatccctgt ccaaaggttgttttttttcccttctat
tgagcttcttgccaccttatcagtttt ttttatgacagtcaaatgacatgatat
atgtgagcatacatggtaatttttaat tctatataaatgaatcactaaataaat
taggaggatatatagtccacctttaag cgtattacacgtgtcacatgaatgtgt
ggcgacttaattgtagaggtttaaatg tagcttcctataatagatgtgttccta
aactacattttaatcattggacttgta tttttatgttagcacttgctgttgaag
aaaagcctatgccaaaagttcagtgaa accaataatccactgccagctttctga
gttaaaaaaaatccctgggttttcaca cacaggaacaccctgtgtgaaacactc
atttagagcaaaatgcatctgataagg agttcctgttgtgcctcaactggttaa
ggacctgacattctccatgagaatgtg agtttgatccccggccccactcgatgg
gttaaggatctggtgttgccacaaact gcagctccgattcatctcctagcctag
aaacttccacagcccagaatatgccac agaattcggctgtttaaaaaaaaaaag
aaaaaaaaaagaatcataaatgtgttg gtttgttcaccaaatacatgataactt
gctcttgccaagctcagcttcataaat attaagtcatttaatacagcagccacc
ttatgaacagatattactatacttccc atttacagataaggaaaatgccatatt
taaccaagagattaaataactttcccg aggtcttatagcaagtaaatcatggtg
caggggtttgaccacacgcagtctatc tccagagtctgtgtatttagccactgt
tttactttcaaatttaaatttataaaa cttctaaattatctgttaaccataatc
tttggaatttttaaaaccacgagttcc tataaaatgtttcattgaaagtaagtc
acttttccatagcttttgataatacat ctgtaggataaaagccacagctctctt
gcagacttggtacaccctggggcaaag catcatgcctgtcacgtacatggtggt
ccttactttgactctcagtgcttttat tgcccaggaattttgtgagatttctag
ttgttgaggtttgtttaaagaggttat gccggtacttggaagagctcttttctt
gctacctggagccttctcatatttcct ttttgaggagggacatgaattgccttt
caaactcataaatatattttctagtac acaagtctccatcttccttagacgcat
ggctcctggagttctccatcctcctgc tccactttgggtgggctcctctctggg
tctgccaccaatctgccacccagagac atccttgacccacttccagaccccacc
atggcttcactttcttcgctttcctcc tttgtggaaccttctgcttaagaatct
gaggaagaaaatttgcacgtgagctaa actggaggtactttcctgcctggtctt
gcacgatagcttggctgagcccatgat gctgggtggctgttactttccatggac
acccgaaggcgttgctcctttggcttc tagttgcatgcagtgttgcttatccca
ggctgatctttcttccactgtaggtga cttttaagaattaagggattaatctat
atctacaacaacaacaacaaagacctt ttcaagctgaggtagggctttctgtat
atgtttggagtggttatccagcagact ttacttgaaggcaggggtcatatcctc
aagtgctcataaacggaccacagaaag atctcataattgggtggagctgggtgg
ggaccgtgtcatgtggccaggaaatgc cagatgggaagggagtggcccttactg
agctccagctgaactctgaattttcta gaaaactcagaaatctggatttttcat
gtgtaatacccagatttatagatgtgg aaagctaattctttttttttttaaggg
actataggcaatgaactaagatctagg ttgtatttggacaaggggtcatcagtt
taagctgtgtagttgagcgctcagcta ttgggctgagggacccctaaatactga
gacggggaggtccttgctctggggcat cacaagtacactccctggtctcattca
aacacttttcctacaaaattgatccca tttcttcagtgcactgtctgaatgcat
ttggcccagagccgtgctgaggcatag ggaaggggtccacggtttcatggcatc
gttttgtgctgtgtgtccctgctgtcg tccaggatacctacctctcctcctcct
gcatctgaatgtccccccacagactct ctgggattctacagcctctggcctgtt
cctcagacacctcttacctgccagctt tccagattcacattagttagtccaaat
ctactgccgtcagtgactcacttcatt tcttcttctccgaggcagttcagcccg
gtacagttgttttgtcaacacttcagt tgagtctggaagatgtgcatgggttat
gcacgagagcggtccatcattttgagc tagaagtcctttctcagcccagagaca
agtcctcatctcctttacttcctgact cttcttcctctgcatccttccaagata
tctctttctccagccaccacctaaatc tcttcttttcccggggttccgtgctca
acccactcttcttcttaaatctgtggc tgggtgaacgcatctgctggcaccact
tctctgctaaagactccaaaaatccat aggtcctgcccggcctttgcccacctc
tctccaacactgtccagctttagatgt agagctaatccccccagagatatcatt
ccctggatgtctaagtcctttggtatc tcactttcagcgtgttcaaaatcctct
tacaactgttctttctccttttccatc ttgattattggcaacatgccagccttt
cccctacccccagcagtgagccaagct agaaacaagggcttaatcttcaatctt
tccttctccatccctaaacctaatgag tctccaagcccttcccagtttacaccc
taaatgttgctcaaaacatcccctagt tcttccacgtgctctcctctatattga
aaggtcaagaaaggccatcttccctcc actgtgaggaaatagatcttgatactg
cccctgagctgggcagtcctcgacctg acaaactgtgcagtgtttctaaatctc
tactggcaaaatgagagtgcctttgac ctgtgttgcgatctcagatcacagtgg
atgtaattgttttataggaatggtgaa cgaaaaagaagtaaatccctaatgcca
aactcctgatcattctatgtcatttaa tagcctgtcatttatgataaagtttcc
tctactggcattagcacaatacttctc aggaaaaaaaaatatgatgccagatac
tgaaaagctcctgggtaaacatgaaca tgggtaccgataaaatggtgaagccag
tccaatcttagagtgacttcccttcat gctacttcatgctcttttttttttttt
tttttaagaaaaacccctttttttttt ctcacaccagtcacagaggagaccgag
gcttagcaaggttaaggtcacatgatt agtaagtgctgggctgaaactcaaaac
catctctgcttgtctcctaaccctgtg cacctctgactattcaacag
ATCCTGTGTCAGGAGTTGGGATTCTTT Exon 15a Seq ID No 25 GAAG
gtaagggccttgaccaccgaattaagg Intron Seq ID No 43
taatcttgctctgtggcaggccttgtt 15a ttcagtattttaagtacactggctcag
gtaatcctcacaacagccccaggagga atgttctattacctccactgtatagat
gaggaacttgaggcacagaatggttgc caaggtcacacagctatattgggggtt
catacccagccatccaactctgtctgt actctctgccactctgcacccccagct
cctgatccacttcctgtttccatccct cgatttctgctgcactcaggggcccct
ctccccctcggcctgtgagatctgctt cagtaggcttttctccctgactcctcc
atccctgtccttacaggcagctgcttc tctccgggacacgaggggtccatacgg
acactctctactggctgggttgcgcct aactcgtgattcctcctctgtttcag
ATTCGGAGCCGGGTTGATGTCATCAGA Exon 16 Seq ID No 26
CACGTGGTAAAGAATGGTCTGCTCTGG GATGACTTGTACATAGGATTCCAAACC
CGGCTTCAGCGGGATCCTGATATATAC CATCATCT gtaagtccgaaaatgcctgtcgtgtgt
Intron Seq ID No 44 gccttaggctgctgcggaggaggccag 16
ggctatataagcagagtcagtgactga ctgtgccctgcagtgttgatggccatg
gagattccaccgttagagcttttttct ttgttaaccttgaaggcaaatctggtt
aggaagataactttcaaagagtcacca tctggacattcatgcccatgtgcttca
atcctgtatacaagcagtttagagtac agggaagggaaggacattatgaaaggg
agagggtgtgtttggatccagcagctc catcctcagaatttatctgaagacact
gcaaaattactaagaatcactatgaca agaatgaggatggggtgatatggcaaa
gttgtgatcctggaagaccttcatctc ccatgttgcccaactctgaacatgaat
ttggtgaactagttggttaaggggatg atcctccaagtttctccctggttgagc
tccaaaaaccatgtaagtttctcatag caaaaccgtataggtccttagggcttt
agttggaatatttgtgctgaaatgctg gaaagccccatttgccatttttgtatt
tgcaaaataatcatcaagaggggagaa tgcattctttcatgaccactgaccctc
tgaaaaggtcaggaatttagtctgaag taggcaagcctcctaccccgcttctgc
catgagcttgcacgcacaggcctgtct tgacatttcttctttatagatttcttt
ttgaatatcttgaaattgctttaaaaa tatttaaagaatgtagaattatataaa
ataaaaaggaaataaccccacacctcc cacaaaaccctgtttcctgcctttctc
cacccactctccagggtaacacttggt aacagcatagttgtatcaccccaggcc
tatttttgagcatatcagcatttcaag aaatgtattttttctcaataaaacatc
ccttatagttgaggaggggaggttatc attcctgggttugtttttttttttttt
ttaatgtaatcctggtacatcggtaat ttgcattttttattcattaatatcttt
ggtatttctagtgttgggacacacagg tcaacctcagtttttgggttttttttt
ttgtctttttgtctttctagggccaca cctgcagcatatggacgttcccaagct
aggagtctaatcagagctgtagccacc agcctacgtcatagccatagcaacgtc
agatccaagccgtgtctgtgacctaca agcacagctcatggcaacaccggatcc
ttaaccactgaacgaggccaggggatc gaacacacatcctcatggatcctagtc
atgttcattaaccactgagtcatgatg ggaactccaacttcaactattttaatg
tctgtaaaacattccatttggaaacca tttcatttgtaaagcaaaatgaaaaca
ttttgttcattttcaacagagttcgta gctgacttctgttctggaaaaaaggaa
atggagcaaatttgagtgagaaagatt caaagataacttttcttttaaaaaaaa
ttatatcttggaaacttctgggctatt gattctgaagactatttttctatatac
tgttttgatagcaaagttcataaatgt gaaaggatcctgcgatgaatcttggga
agcagtcatagcccaatatatctttgt tgcttttaaaatgagatttagtttact
aaatatttttctgatcataaaaataac acagatctaccgcagaaaatttggaaa
aaaaaaaacttttaaattcaaaaaaca gttaaaccacaaatgatcccaccatcc
agagagcaatttgtactttggtgtcta gttcatctttctttttctgtttacaag
cacatataccacaagcattttttcaaa aaatgaaaatgggataatactatacat
acgtctgtacacctgcatagttactga acagtctttgatctaccctgtaagttt
ctaacttttcattatttgaaatgatgt tttggcaaagaaatatgtaggtgtgtc
tcgcacactttcataatgatttcttag gataaatttcttaggataaattcataa
tgatttcttataataatccatactctg ccaactgatcttcagggaagccaactc
gccttctcagaaataacatataaccca tttacttgccctctcaccaatactagg
tcctaatgtttttgtgtacagattcta tatttttacatacaagaattccttaaa
gcaaggcatgtcacagaaaaatagaag gaagacacaattgtcatgtttaaggac
tgcattctgtaccaaaaatgctaagtt aaatgaacatctgaaacagtacagaaa
cgctatctttcagggaaagctgagtac caggtactgaacagattttggcaaata
cagcaggcatggatgtttccaaaacat gtttttctactttatctcttacag
GTTTTGGAATCATTTTCAAATAAAACT Exon 17 Seq ID No 27
CCCCCTCACACCACCTGACTGGAAGTC CTTCCTGATGTGCTCTGGGTAGAGAGG
ACCTGAGCTGTCCCAG gtaaagcatcctgcaggtctgggagac Intron Seq ID No 45
actcttattctccagcccatcacactg 17 tgtttggcatcagaattaagcaggcac
tatgcctatcagaaaacctgacttttg ggggaatgaaagaagctaacattacaa
gaatgtctgtgtttaaaaataagtcaa taagggagttcccatcgtggctcagtg
gtaacgaaccctactagtatccattga ggacacaggttcaatatctggcctcac
tcagtcggctaaggatccagtgatgcc gtgagctgcagtgtaggccacagacgt
ggctcagatctggtgctgctgtggcta tggtgtaggccggccccctgtaactcc
aattcgacccctaggctgggaacctaa aaagaccccaaaaaagtcgcttaatga
atagtgaatacatccagcccaaagtcc acagactctttggtctggttgtggcaa
acatacagccagttaacaaacaagaca aaaattatcctaggtggtcagtggggg
ttcagagctgaatcctgaacactggaa ggaaaacagcaaccaaatccaaatact
gtatggttttgcttatatgtagaatct aaattcaaagcaaatgagcaaaccaat
tgaaacagttatggaagacaagcaggt ggttgtcaggggggagataaggggagg
caggaaagacctgggcgagggagatta agaggtaccaactttcagttgcaaaac
aaatgagtcaccagtatgaaatgtgca atgtgggaaatacaggccataacttta
taatctcttttttttttttgtcttttt tgccttttctaaggctgctcccgtggc
atatggaggttcccaggctaggagtcc aaacagagctgtagctgccagcctaca
ccagagccacagcaacacgggaacctt aacccgctgagcaaggccagggatcga
acccgagtcctcacagatgccagtagg gttcattaaccactgagccacgacagg
aattccagggtctgttgtgttcttaaa acacttccaggagagtgagtggtatgt
cataagtaaacaataaatgttaaccac aacaagcttatgaaataaacaggaaag
ccatatgacctacaatcagtcattggg agaatccacaaaaggttgagcagagga
tcaattccagctcacactccagtttta gattctcccctgccttaaagcatcaca
gactacataatctgagctgaagaataa aaattaaaactcaccccagtgcaaaac
agaaatgaaaaagtattaaaacgaggt tcatactgttgttcattagcaatatct tttattcacag
GGGTGCCCAACAACATGAAAAAATCAA Exon 18 Seq ID No 28
GAATTTATTGCTGCTACGTCAAAGCTT ATACCAGAGATTATGCCTTATAGACAT
TAGCAATGGATAATTATATGTTGCACT TGTGAAATGTGCACATATCCTGTTTAT
GAATCACCACATAGCCAGATTATCAAT ATTTTACTTATTTCGTAAAAAATCCAC
AATTTTCCATAACAGAATCAACGTGTG CAATAGGAACAAGATTGCTATGGAAAA
CGAGGGTAACAGGAGGAGATATTAATC CAAGCATAGAAGAAATAGACAAATGAG
GGGCCATAAGGGGAATATAGGGAAGAG AAAAAAATTAAGATGGAATTTTAAAAG
GAGAATGTAAAAAATAGATATTTGTTC CTTAATAGGTTGATTCCTCAAATAGAG
CCCATGAATATAATCAAATAGGAAGGG TTCATGACTGTTTTCAATTTTTCAAAA
AGCTTTGTTGAAATCATAGACTTGCAA AACAAGGCTGTAGAGGCCACCCTAAAA
TGGAAAATTTCACTGGGACTGAAATTA TTTTGATTCAATGACAAAATTTGTTAT
TTACTGCGGATTATAAACTCTAACAAA TAGCGATCTCTTTGCTTCATAAAAACA
TAAACACTAGCTAGTAATAAAATGAGT TCTGCAG
[0089]
11TABLE 10 Genomic Sequence of CMP-Neu5Ac Hydroxylase gene Seq ID
No. 46
ctgccagcctaagccacagccacagcaacgctgggtctgagccatgtctgcagcctatgccagagctccccgc-
agcg ccggatgcttaacccactgagcaaggccagggattgaaccctcgtcctcatg-
gatagcagttgagttgtttccacggaa ctcttaggggaactcctgattattttttat-
ttaaatttatatttctctgactttttcgtgtgctcatcagccactgactgtgtatctcc
attagtcatggtttgttaactctgtcattcaaaccctcttcatccttgctacgcagataacatcattataat-
aaaatcgtgcctg aagaccagtgacgcccccaagctaagttactgcttcccctgggg-
ggaaaaagaagcaccgcgcgggcgctgacacg aagtccgggcagaggaagacggggc-
agaggaagacgggggagcagtgggagcagcgggcagggcgcgggaa
gcactggggatgttccgcgttggcaggagggtgttgggcgagctcccggtgatgcaggggggaggagcctttt-
ccga agtagcgggacaagagccacgggaaggaactgttctgagttcccagtCCCGAC-
GTCCTGGCAGCGC CCAGGCACTGTTATTGGTGCCTCCTGTGTCCACGCGCTTCCCGG- CCAGG
CAGCCCTGGCGGATCCTATTTTCTGTTCCCCCGATTCTGGTACCTCTCCC
TCCCGCCCTCGGTGCGCAGCCGTCCTCCTGCAGTGCCTGCTCCTCCAGG
GGCGAAACCGATCAGGGATCAGGCCACCCGCCTCCTGAACATCCCTCCT
TAGTTCCCACAGgtgagaaggcttcgccgctgctgccgctggcgccggcagcgccctccacgcacttcgt
agtgggcgcgcgccctcctgcattgtttctaaaagatttttttttatccgcttatgctat-
cagttactgaggaagtatttacaaa tctactattattttgaatttgcctttttctcc-
ttatagtttatcagtatctcttgagactgttattggtgcctgcaaatttaaaatgatt
ggggttttatgaggaagtgaaccttttatctttatgaaacgcctaactgaggcaatgttaattgcttaaaata-
ctttctttattat cagtgtggccatgccagtgtcctcttggttagaatttgcctgat-
.........................................ctgcc
aaagctgggagatgggggaaagtagagtgggttattgaaactgaatatagagttcagcatctaaaagcgaggt-
agtag aggaggaagctgtgtcaacggaaatactgagctgggttcacatcctctttct-
ccacacagTCTAATGCCTTG TGGAAGCAAATGAGCCACAGAAGCTGAAGGAAAAACC-
ACCATTCTTTC TTAATACCTGGAGAGAGGCAACGACAGACTATGAGCAGgcaagtga- gaggggg
ctttagctgtcagggaaggcggagataaacccttgatgggtaggatggcc-
attgaaaggaggggagaaatttgcccca gcaggtagccaccaagcttggggacttgg-
agggagggctttcaaacgtattttcataaaaaagacctgtggagctgtca
atgctcagggattctctcttaaaatctaacagtattaatctgctaaaacatttgccttttcatagCATCGAAC-
AAA CGACGGAGATCCTGTTGTGCCTCTCACCTGCCGAAGCTGCCAATCTCAA
GGAAGGAATCAATTTTGTTCGAAATAAGAGCACTGGCAAGGATTACAT
CTTATTTAAGAATAAGAGCCGCCTGAAGGCATGTAAGAACATGTGCAA
GCACCAAGGAGGCCTCTTCATTAAAGACATTGAGGATCTAAATGGAAGg
tactgagaatcctttgctttctccctggcgatcctttctcccaattaggtttggcaggaaatgtgctcattga-
gaaattttaaat gatccaatcaacatgctatttcccccagcacatgcctaacttttt-
cttaagctcctttacggcagctctctgattttgatttatg
accttgacttaatttcccatcctctctgaagaactattgtttaaaatgtattcctagttgataaacagtgaaa-
cttctaaggcac atgtgtgtgtgtgtgtgtgtgtgtgtgtgtttaccagcttttata-
ttcaaagactcaagcctcttttggatttcctttcctgctctct
cagaagtgtgtgtgtgaggtgagtgcttgtccaaacactgccctagaacagagagactttccctgatgaaaac-
ccgaaa aatggcagagctctagctgcacctggcctcaacagcggctcttctgatcat-
ttcttggaagaacgagtgctggtacccctt ttccccagccccttgattaaacctgca-
tatcgcttgcctccccatctcaggagcaattctaggagggagggtgggctttct
tttcaggattgacaaagctacccagcttgcaaaccagggggatctggggggggggtttgcacctgatgctccc-
ccact gataatgaatgagggattgaccccatcttttcaagctttgcttcagcctaac-
ttgactctcgtagtgtttcagccgtttccata ttaggcttgtcttccaccgtgtcgt-
gtcgtcaatcttatttctcaggtcatctgtgggcagtttagtgcgaatggactcagag
gtaactggtagctgtccaagagctccctgctctaactgtatagaagatcaccacccaagtctggaatcttctt-
acactggc ccacagacttgcatcactgcatacttagcttcagggcccagctcccagg-
ttaagtgctgtcatacctgtagcttgcttggct ctgcagatagggttgctagattag-
gcaaatagagggtgcccagtcaaatttgcatttcagataaacaacgaatatattttta
gttagatatgtttcaggcactgcatgggacatacttttggtaggcagcctactctggaagaacctcttggttg-
tttgctgaca gactgcttttgagtcccttgcatcttctgggtggtttcaagttaggg-
agacctcagccataggttgttctgtcaccaagaag
cttctgcaagcacgtgcaggccttgaggtcttccgacttgtggcccggggactctgctttttctctgtccttt-
tttctccttagt gggccatgtcctgtggtgttgtcttagccagttgtttaagggagt-
gttgcagctttatgattaagagcatggtctttccttgca
aactgcttggtttagaagcctggctccaccacttagcggctctgtgacctcggacacatttcttagcctttct-
gggcctcgc tcttcttcctcataaagtgaaaatgaaagtagacaaagccttctctgt-
ctggctactgagaggatggagtgatttcatacac ataaagcacttaaaataatgtct-
ggcatatgatacatgctcaataaatgtcacttacatttgctattattattactctgccatgat
cttgtgtagcttaagaacagaggtctttacaggaattcaggctgttcttgaatctggcttgctcagct-
taatatggtaattgct ttgccacagactggtcttcctctccttcacccaaagcctta-
gggggtgaacgatcccagtttcaacctattctgttggcagg
ctaacatggagatggcaccatcttagctctgctgcaggtggggagccagattcacccagctttgctcccagat-
acagct ccccaagcatttatatgctgaaactccatcccaagagcagtctacatggta-
cactcccccatccatctctccaaatttggct gcttctacttaggctctctgtgcagc-
aattcacctgaaatatctcttccacgatacagtcaagggcagtgacctacctgttc
caccttcccttcctcagccatttttcttctttgtacataatcaagatcaggaactctcataagctgtggtcct-
cattttgtcaatc taatttcacagcctcttggcacatgaagctgtcctctctctcct-
ttctgcctactgcccatgagcagttgtgacactgccaca
tttctcctttaacgacccagcctgctgaatagctgcatttggaatgttttcaatttttgttaatttatttatt-
tcatcttttttttttttttt ttttttttttttttttagggccgcacccatgggatat-
ggaggttcccaggctagggatccaatgggagctgtagctgctggcc
tacaccacagccacagcaatgcacaattcgagccaatctttgacctacaccagagctcacggcaacactggat-
tcttaa cccactgattgaggccagggatcaaactctcgtcctcatagatacgagtca-
gattcgttaacctctgagccatgatagttg ttagttactcattgatgagaaaggaag-
tgtcacaaaatatcctccataagtcgaagtttgaatatgttttctgccttgttacta
gaaaagagcattaaaaattcttgattggaatgaagcttggaaaaaatcagcatagtttactgatatataagtg-
aaaataga ccttgttagtttaaaccatctgatatttctggtggaagacatatttgtc-
tgtaaaaaaaaaaaatcttgaacctgtttaaaaaaa
aaacttgactggaaacactaccaaaatatgggagttcctactgggacacagcagaaatgaatctaactagtat-
ccatgag gacacaggtttgatgcctggcctcgctaagtgggttaaggatatggtgtt-
gctgcagctccaattcaacccctatcctggg aacccccatatgccaccctaaaaagc-
aaaaagaaaggtgctgccctaaaaagcaaaaagaaagaaagaaagacagc
cagacagactaccaaatatggagaggaaatggaacttttaggccctatctccaactatcacatccctatcacc-
gtctggta agaaatggaaaaaatattactaagcctcctttgttgctacaattaatct-
gattctcattctgaagcagtgttgccagagttaac
aaataaaaatgcaaagctgggtagttaaatttgaattacagataaacaaattttcagtatatgttcaatatcg-
tgtaagacgtt ttaaaataattttttatttatctgaaatttatatttttcctgtatt-
ttatctggcaaccatgatcagaaatctttaaacaatcaggaa
gtcttttttcttagacaaatgaaaatttgagttgatcttaggtttagtacactatactaggggccaagggtta-
tagtgtgactat taaatcacagataatctttattactacattatttccttatactgg-
ccccacttggatcttacccagcttagcttttgtatgagagt
catccttaaagatgactttattctttaaaaaaaaaaacaaattttaagggctgcacccatagcatatagaagt-
tcctaggcta gcggtcaaattagagctgcagctgccagcctatgccacagccacagc-
aatgccagatctgagctgcatctgtgaccta cactgcagcttgcagcaatgctggat-
ccttaacccattgaacaatgccagggattgaacacacatcctcatggatactgc
tcaggttcctaacctgctgagccacagttggaactccaaagcagactttattctgatggctctgctgatctct-
aacacgttat tttgtgccatggtgtttatcttcactttactcaagtcagggaaacac-
gaagagtctcatacaggataaacccaaggagaaa tgtgcaaagtcacatacaaatca-
aactgacaaaaatcaaatacaaggaaaaaatatcttcactttcaaaatcacctactga
tgatgagtttatatttccttggatatttgaatattagctatttttttcctttcatgagttttgtgttcaacca-
actacagtcgtttactt tgatcacagaataatgcatttaagccttaaatagattaat-
atttattttcaccatttcataaacctaagtacaatttccatccag
GTCTGTTAAATGCACAAAACACAACTGGAAGTTAGATGTAAGCAGCAT
GAAGTATATCAATCCTCCTGGAAGCTTCTGTCAAGACGAACTGGgtaaatac
catcaatactgatcaatgttttctgctgttactgtcattggggtccctcttgtcaacttgtttccaatctcat-
tagaagccttgga tgcattctgattttaaactgaggtattttaaaagtaaccatcac-
tgaaaattctaggcaagttttctctaaaaaatcccttcattc
attcatttgttcagtaagtatttgatgagaccttaccatgtgtaaacattgcactaggtattaagaaatacaa-
agatggataa gatagagtcggcgtaaatgagatgatataatgagacgttataatgaa-
actcacaattccagttgggaaataaagtccttca aattccatgactctttctggcac-
acgttagaggctacagcttctgtgtgattctcatgctggctccacttccactttttccttctt
cctactcaagaaagcctatagaaatatgagtaagaagggcttaatcataggaataaatttgtctctgt-
tctaagtgattaaa aatgtctttatcagtataaaaagttacttgggaagattcttaa-
aactgcttttacacactgttctagaatgactgttatataaata
aaaaagtagatttgatctaacacaattaaatgacctttggaaatattgactaattctcaccttgcccctcaaa-
gggatgcctg aaccatttccttcttttgccagaaagcccccaccctttgtctgttga-
cctagcctaggaaatcttcagatcacgttgttagca
cgaactggttacatgtgctgtacaaatactatttaattcatctgattaaaaaaaaagagataagaagcaaaag-
tttgactatc ttaaactgtttgcgtaggtgagaggacaattgaccatctactttatg-
agtatgtaacccagaaacttaaagctccttaaggg
agctaagtcttttggataagacctatagtgagaccttttagcaaaatggttaagactgaatggagctcactag-
cgtgggttc atatcctgatgctcaaacacgcaattaaatgactttaggtgggttagt-
ctctgttccttagtttcctcaatgggagataatatt
ggtagtagcgattttactgggttgttgaaagaacatctgttaaatgttcagaacgtgttacgacagagtacag-
agtaatgat ttgcttgtatatgtatgactcaaatagtctgccatatgccttgtgact-
gggtcctgtggagcaggaaggagggatttcccac ccagcagaaagttgggtaaactg-
gaaaatagactgaggccaggaaatgatgcaaagcgttgatgttcactgccacggc
aggtgaagggcagggccagagttgtcagtagggtcaggggaggactggaaataaccaagacccactgcacttt-
tcag cctttgctccagtaaggtaatgttgtgagagtagaaaattttgttaacagaac-
ccacttttcagtacagtgctaccaatactg tagtgatttcataccacatcccaagaa-
agaaaaagatggctcaatcccatgtgagctgagattatttggttttattgttaaat
aaatagcattgtgtggtcatcattaaaaaaggtagatgttaggaaagtagaaggaagaagactctcacctaca-
ttttcatc actgttttggtatctgccagttgtcaccttggtccccttccccgcctct-
cccctgcctcctcttcctccttctcctttttttggaat
acaattcaggtaccataaaatttacccttttagagtgtttgactcaatggtttttagtattttcacatgttgt-
gctattactatcact atataattccaggtcattcacatcaccccccaaagaaacctt-
ctaactattagcagtccattcccttcttccctcagcccctg
gcaaccactaatctacttactgtctccatggatgttcctatattgaatcaagctagcataaaccccacttgct-
catggtcata attcttttttatagtgctaaattacatttgctaatattcaattaagg-
atttctatgtccatattcataaggaatattggtgtgtagttt
tctctttgtgtgatatctttgtctggttgggggatcagagtaataattactgctctcatagaatgaattgaga-
agtgttccctcc ttttctatttattggaagagtttgtgaagtatattggtattgatt-
cttctttaaacatttggtcagattcaccagtgaagccatctg
ggccatggctaatctttgtgaaaagttttttgattactaattaaatctctttaatttgttatgggtctgctcc-
tcagacgttctagt tcttcttgagtcagttttgttcatttgtttcttcctaggactt-
tctccctttcatttggattatttagattgatagtaatatccccctttt
aattcctggctgtagtaatttgggtcttttctcttttttcttggtcagtttagctaaaggtttgtaattgtat-
taatcttttcaaataa ctaacttttttgttttgtttgttttttgttttttgtttttt-
gttttttgtttttttttgctttttaaggctgcacctgaggcatatggaagttct
caggctagaggtctaatcggagctacagctgctggcctataccacaaccatagcaatgccagattcaagctgc-
atctgc gacctacaccacaactcggccagggatcacacccgcaacctcatggttcct-
agtcggatttgttaaccactgtgccacg acgggaactcccgcccattttttttaaca-
cctcatactttaacataaagatgggcttcacatggactgatagctcaaatgag
gaaggtaagactatgaaagtaatggaagaaatgtagactatttttgtgacctagagattactgatacttcttg-
acttttcaaa caatacttcaaaagtacagcccaaagggaaaaaagaaagaaaaaaga-
aacacacatatacacaaacctagtgaataa gatatcatcgatacactacagatttct-
atgaactggaagaccccatggacaaagttaaagaacatatgatagtttgagtga
ttattttgcaatatttacaaccaatgagggaatattatccagcttataggaggaagtaatgcaaatcgacaag-
aaaaagata ggaaacccaatataaaaattaagaaaatacaaaaattaagaaaggata-
tgaactagcattttacaaaagaaaaatctcca aaagtcaatcagcacatgaaaatat-
gctcaaacctattaattattagaaaactacagactgaagcaatgaggtgctttactt
tacatctttttgactgataaaaagttagaaacaaaggtgatatcaaatgtcagggataaaaggatatagaaat-
cgtcatgc ctgtggtgggagtatggccggtgcagtcatgtgggaaggtaatctgaca-
gtggttaggcagagcaggtttatgaataca ctgtggcccatcaatcccacgcctgtt-
tatgtaccaaagaaatcctgttgtggcagaatctatgggtccacccctgggag
catgaattaataaaatgtggcaccagggtgtgtgaaactccagctagagatgagatgtccacatggcaacatg-
aatgca tcttagaaacatagatttgagtgaaaaagagtaagaaacagccgggaaacc-
caataccatttataaaaattaaagatgca cacatacaatgtagtaaatattttgcat-
gaactttcaaatggttgcctacagggggggagagtaaagaagagtagaaaac
aaagataaagggagtaagtaagtagctctgcctggactgaatataatgtgtcatgaactgagaaatatggtta-
acataatc ctcttaacttgaggtcctaaatgaatgaatgagtccactattcatttac-
ccattctttaatgtgtattgcattataatccattttttt
agaaccaacgaattttgttcccataactactaatcagcctgccttttctccctcattcccttatcagctcagg-
ggcattcctag tttttcaaacgttcctcatttgaaccaaaaatagcatcattgttta-
aattatacttgttttcaaatacgatgcttatatattccaagt
gtgtttgcccattttcttaggtggtagaaatttttcattctacttttctatctactcagattttcccgttgga-
attatttccattgctat taaacttagaagtcccccctgtgatatgccatttttttca-
tactttttaagcacttggttgcttttctttgtgtctttaagcacctag
aatacttataaccattgcacagcactgtgtatcaggcagcccttcctcttccactaatttatggtccttctct-
tagactatatta aactgttatttaattaggatcctctcttcgtccttatgatttaat-
tattatagttttctaatatgtttttattataattcctcttcattattc
ctccctattaaaaattttaatgaattccatttgtttgttcttctagttaaatattaagtcataatccaaataa-
cttagatgtcattag tttatgtggtcaaagtaaggataccacatctttatagatgca-
ggcagttggcagatgtcatgattttcttcagtgcataaatg
caatttatctttgagcaaggggcataaaaacttttatggtattggctttgaaataatagttaagaactgcaga-
ctcagtttttc ctgcttttcttgaaaaagaacacttctaaagaaggaaaatccttaa-
gcatggatatcgatgtaattttctgaaagtctcctgta
attccttgggatttttgttgttgtttgttggtcggtttttttgggtttttgtttgtttgttttgttttgtttt-
gttttgcttttagggctgcac ctgtggcatatggaagttcccaggctaggggtccaa-
ctggagctacagctgccagcctactccacagccacagcaaca
tgggatcctagctgcatctgtgacctaaccacagctcttggtaatgccagattgttaacccactgagcaatgc-
cagagat cgaatctgcctcctcatggacactagtcagattagtttctgctgagccac-
aatgggaattcccaattccttgtatttttgaact ggttatgtgctagcatataattt-
tgtttcttgaatctttgtgggtttttttttttttttttttttgtctcttgtctttttaaggctgcaccca
cagcatatggaggttcccaggctagaggtcaaattggagctacagctgccagcctacacaa-
caactgcagcaaagtg gggcccaacttatatgacagttcgtggcaatgccggattcc-
taacccactgagcagggccagggatcgaacctgagttt
ccagtcagtttcgttaaccactgagccatgatagtaactcctgtttgttcagtcttgaacctcctttttaatt-
ctttattccttgag ggtgaaataattgccataataatactatcatttattacatgcc-
ttctctgtgctaggcatagtgacactttaggatttattatatc
acttaatccctacaacaactctgcaaagtatgtatcataatcctatttgacagatcaggaaattgcagcccag-
gatgcaga taatatgcatccatcacaagtgactagatatagtccctctgctattcag-
cagggtctcattgcctttccattccaaatgcaat agtttgcatctattgtatatgtg-
ttttggggtttttttgtctttttttttttttttgtcttttctggggcctcacccttggcataggtagg
ttcccaggctaggggtcaaattgaagctgcagctgccagcctacaccacagccacagcaactc-
gggatctgagcctca tctgcaacctacaccaaagctcacggcaacaccggatcctta-
acccactgagtgaggccagagatcaaaccggcaac
ctcatggttcctagtcggattcattaaccactgagccacgatgggaactccctaaatgcaatagtttgctcta-
ttaacccca aactcccagtccatcccactccctcctcctccctcttggcaaccacaa-
gtctgttctccatgtccatgattttcttttctgggg
aaagtttcattttgccatttttcattttacgggtaatttttacttcagtttcttccactagcagttgtcttaa-
agtgagtataattaa tattcatttggaaaatgtaagcaaaacattttttaaagggcc-
atgcccacagcatatgaaagtttctgggccaggggttga
atccaggctccaagttgcagctgtgccctacactgcagctgggcaatgctggatcctttaacccactgtgccc-
ggctag ggatcaaacctgcatttccacagctacccgagccattgcagttggattctt-
aacccactgcactacagtgggaactccca caaaacattttttaatgtcctttgaata-
aagtaggaaagtgctcgtctttgagggcagggcggcaatgccatttccacaag
gtttgctttggcttgggacctcatctgctgtcatttagtaatgaataaaattgctgacagtaataggattaac-
tgtgtgtggag atagccagggttagagataaaaacactggagaagtcaaataagttg-
ctcgaggtcctctagctaataagctattaagtgg gagagtgagggctagaaacaggc-
catctgtctcccaagcacatgtccattagtggtttgctgatagccttccagaacaa
cagagaggactctcaaacatggtcttgcctccctccaattgatcccctccatgtgcctcacagcgggtctttc-
taaaattaa gttctgattttaattctcccttgctatagcacttaggtatggctttca-
gccgtgcaataaaaagcaggcaagagtggctcaat
catataggaggttgtttttcttagatcccaagcaggtaatcctgggcattatggttgttctgcgtttatcaag-
gagccaaattc tctatcacctcctgttctatcctcctcagtatctggctctattctt-
cagcatctcaagatggcttgtgctcctccaagcatggc
agtcaaattccacacaagagggggaaatatgaagggcagacagtgctggtctcctgagctgtccctctttgtc-
ggggaa ataaatgtattccttcatcccgtgagacttctgaagtagacgtctgcttac-
gtctcacccaccagaactatgtaaactgc acatagtgctaggtctacatagccactc-
ataactgccagggggtgggaaatctttaaataggtgtaccaccacacaatta
ggatgctaatagtaagggagaaggagagaataggttttgcgcaagccaccagcatgcctgccacaattgctta-
aaattc ttcattgacccctcattgccacaggatgaaatccaaacgccttcttagttg-
ggaatctgacctacctgtctctcccacctgg ttcagacaccattctccttggtcata-
aaattccagtcatttgtgaacatccagctcccccatgcctccatgcctttgcacatg
ctgttcttttatcttttatgttgtccttttatcttttatccaaaagagatatcccatcatcacatctcttttg-
tcagcccccaaatact ttgtctttcaagttcagctggaggattacctcctatttgaa-
atcagctttgtctcttacaaccaaacaaggttttccttccgaga
cactcccacagcaccttgaactcatctctatcaatcattcatttgattgtaatgaagttgttggtggtatgcc-
tgtgtctctgac acatctgcgatctcatgagttccttaagtggaatgtgaatagcgg-
gatgaacagtattggtcttcagccctcatctctgcag
atgttgcttgacccaaatgagcgttgccttttattttgattttgctttgatttgtctactccatgtacttgag-
ccatgcatttctgtc ttagcgatgctttttaaaagtcattttttggttgattatcca-
gatttgtccacctttgcttctagTTGTAGAAAAGGA
TGAAGAAAATGGAGTTTTGCTTCTAGAACTAAATCCTCCTAACCCGTGG
GATTCAGAACCCAGATCTCCTGAAGATTTGGCATTTGGGGAAGTGCAGg
taaggaaatgttaaattgcaatattcttaaaaacacaaataaagctaacatatcaatttatatatatatatat-
atatatatttttttt tttttttacatcttatattaccttgagtattcttggaagtgg-
ctagttaggacatataataaagttattctgaagtctttttttttctttt
tccatggtgagcagtggcttgatgtggatctcagctcccagacgaggcactgaacctgagccgcagtggtgaa-
agcac caagttctagccactagaccaccagggaactccctattctaaattcttgagc-
acattatttaggaacctcaggaacttggc aggattacaggaaatatatctagattta-
aaaaaaaatcttttaacagaggtcccaaaggagagtcatgcacagctatggg
aggaagttcagaaactgcccttgctaccagatcactgtcagataaaatggccagctacatgtttctgcacatt-
gccctaag atctttacaaacttttctgtgcatttttccacttttaaaagaaaatttc-
ggggttcctgttgttgctcagtggttaacgaacccaa
ctagtatccatggggacaggggttcgagccctggcctcactcagtgggttaagaatctggcattgctgtggct-
gtggcgt aggctggcggctacagctcagattggacccctagcctgagaacctccata-
tgccgcaggtatggccctaaaaaaaaaa aaaaagagagagagagaatttcctccaga-
aaaaacactttggtagtttgggagaagtaaacaaccaaaaattaatttttct
ggagtattcgggaagcttgtaaaaatgggctcttacttttttgaggagacaaatgggaacctacccagaagag-
gcacaat cacctgcatttgatttcttgacctctccctaccttctttgctggcfficc-
acatttggatttctgtgaccttatctctgctccttggt
gttttcatttttcctgtggacgtgccagactatgggaagggagtaaggcgttgatttagaatcctgtagtctc-
tgcctgtctct agtcattgttttcacccttctcaaaggaccttgacatcctgagtga-
gtccgcaagtaatttaggggagaagccttagaagc
cagtgcagccaggctacatgactgtgtccacccactggaaccagtcatttttatacctattcacagcccccct-
accatttaa atccccagaggtctgccataacatctgtaactccctttcctggtaaat-
tgtgttctaaaagactggtaacaaaagatattctg
tggtacagagcataattaaatacctgggagctgatttgagtggggtaaatcaactggtttgacccctaaaacc-
caccatg agcatttctgttctaataaagtaatgcccgtgctgggaattgtgttctac-
ggaaatgctcctgctgtgtctttcttgagtcctgt
gtcattgaacatgcttaggagcaaaggtcccccatgtggctgtctgctaaccagcccagttccttgttctggc-
tggtaatg atccgatcatctgaatctcactgtcttccaacagATCACGTACCTTACT-
CACGCCTGCATGGA CCTCAAGCTGGGGGACAAGAGAATGGTGTTCGACCCTTGGTTA- ATCGGT
CCTGCTTTTGCGCGAGGATGGTGGTTACTACACGAGCCTCCATCTGATT
GGCTGGAGAGGCTGAGCCGCGCAGACTTAATTTACATCAGTCACATGC
ACTCAGACCACCTGAGgtaaggaagggtgagccctcaactccgaagaaaatgctgcaataaaagcact
gttggttttcagctttttttgtaatcactgctcattctgaggtagattcgcttgggc-
tgataaaaagagaactaattcagataaa tgcttgcatttgcatagcctcttttttta-
aaaactttttttttttttttttttttttggcttttcagggctgaacctgtggcatatggagg
ttcccaggctaggggtcgaatcagagctgtagccccgggcctatgccactgccatagcaacatgcatag-
cctccttttta aagtgccttcctgttttataccattgggatgtgagaagagctattg-
tggaaangagcatggggtnataaccctggacctct
cacgtcctaccctcaggntagtgggaaaactctgagtttaaggacatcaaagtgactcctttttagttacatt-
atggngga atcagcncatatttttacaaggggcggagngtaanctgttggagtttac-
aagacatatggtggcattgcaactacttaacc ctactattatagcacaaaagcagcc-
atagtcggtcctgaaggagcctgatgccttcagctttataggcaatgacgtgtga
atatcacaaacagtttcctgtgtcaccaaacatgattgccttttgatttccctttcaaccctttaaaaaaagg-
taaaagccctt cttagcattcagcagcaggtcgctgtgttttgccaactcctgatct-
gtagcatttcgacaacactgagctctcaacttttgaa
ccctgagtccaccacatccttcagtgaaaccagagccatgtgatactaaggatagaaacggaaacttcctgaa-
tccagg cgatcaaataggagggagaaagaggaactttcattgacaaaaccacaaata-
ttgtgaatggactgttacaaatattgtga atgctcctattcccaaccccctggcttc-
attacagggtcctatgtgttcatccttattgagaaatttgtattgctactgccaggt
tgccaatacccagcggtgcccatggtgttctaaaatgaagcaatttcaactttatttttttttcctgtgactt-
tacatgacaagt tcacatgaaggatatactttgatagtaatgtccatggttagggaa-
tatacattgtttgctggttgactggcccctggatttttct
attgaaagtccatgagatctcgaaggcacaggtgtgttctctcgctttttaaggaaagggtttaaaaacttaa-
gtaattaac agctttagtaacaaattacctataacacacttaaaaaccgaataccac-
ccactggagtattgtgctacgattaaaaatctac ttgtctactacatgatatctttg-
tcccacagaaggttctggaaccaaacttgtaatttcaggattatgagagccctgagttca
cgcattgtgtaataactatgttgtgtggtagtcaatttgtacagcttgcttagagagaacaatgtcaagtt-
aaggaggcgatt gctttatagtgcctgtcacaagatgccattgccattgtcctagca-
agagatattctatgggagtatactacattttagtgagg
ataagaactttttatggcatttagtccggtcatttcccaaccactgtcctgaaaaccaatttcattttgattt-
caggggcttgtg tgggcaaagttgccaggcattaaaaagccacttctcaactgtagt-
atcacaatgctttagttgggtagtgtattgcagatag
cttatggctgaaaagttaccaagccttgcagttttcactcctttgagtttatttccttgacagaattgaccct-
gagttttttgact cttacctgctcaactaataaacaccagagtcatttatctccatt-
gctcttgtctgacctttatttaccgaataatgccttatggg
ttcacaaaaacaaggggggagggggccagcatgccttagaaactgtctttagtcaagaaatgngattttatta-
tgtaaat atatgagtattataatagatagtgttattaatagacaccagcaagaattg-
tcaataatttaaaaatcacaaattaaaatacatc catgttagnatcatttatcctaa-
ctcccaaagccctttaaagtggaagatttagatgttaacccagagattaaagacatgttc
aaagaatccttgatttttttttgaatcccttgtttttagagaagaaaacctaatgattttccccctctgga-
ttctacatattaaata tagttttggaacttgaatattagtatggttaataagtgctg-
atatgctgattttgtttatatttttcttatgagtaaatatcctatatc
accagacattatagtctatgtacaaatatgattcttaaacctgatagcacattcattagagttggaattgcct-
tttttttttttttttt tacagttgcacctgcaacatatgaaagttcccaggctaggg-
gttgaatccaagctgcagctgccaccctacattacagc
cgtagtaacagcagatccgagctgcatctgcaacctatgctgcagctcagggcaatgccagatccactgagtg-
aagcc agggatggaacttgcatcctcatagagacaacgtcgtgtccttaacccactg-
agccagaacaggaactccagaatttcc tttcaatagaagaagcaccaagtttaggat-
cagaaagcctgaatttgaataccaatttactatttgttagtcatatatttctga
gtgtgtttcctcatttattaaaagcagactaaaagatgagagggtcttttgttgagaatcaaatacaataaca-
tgtgaaagtg tgtaacactatgattgaaatatacctacacagccatttatttgttta-
ttgttcatgttttgccacccacacagtagtatataatcc
ttttatgtaataaatgctaataatgaaagttggcaacttatgtaagtactcaaaatgctggaggtcatgggat-
actgactgg gatactacagaggtaatgtcatttcctctgcgctaaacttattgtctg-
tagttagggactgactctctttaggacaaggagtt
cattctgtataccatgtgtggctatcacccttcgaagttgaaaaactgccccagggtgggcacccatccgttc-
tcttagata tatggccgagacctttctctcactgggagggaaccacactgaggaatg-
agaaaaaaaaaaggaaaatcaagatgaaa ccagaaacctctttggcataacttctcc-
actctgtactttttgttagaactacccttgcacaaagcagcatcagtgtggaag
acagaatttgcacacctggtttgatatacatgccgtggtatatgggatgttctaacaataaagaggactctcc-
caggaaat ctcctcactgttatagtcagccttgaggaaagagctcttcttttggact-
ctggggagagtctagtttttcagttccttgcttctc
ggtcaacgtgttggtgtaaggatcacactctctcttatactagataattctattttttcacctttcaacctgt-
ctatccttctgac cctagTTACCCAACACTGAAGAAGCTTGCTGAGAGAAGACCAGA- TGTTCC
CATTTATGTTGGCAACACGGAAAGACCTGTATTTTGGAATCTGAATCAG
AGTGGCGTCCAGTTGACTAATATCAATGTAGTGCCATTTGGAATATGGC
AGCAGgtctgtgttctttccacatgtttgggttatcctttctgggataaatttgaggcgagataga-
aactttaagactaaa gaaacaatggcctactttttttgtacatggtcctgtgtaaat-
ctctatttgagctgaaataagatggtcttcctctccaattatc
catggtatgactctgatggataacaaatccagttctgaaaaaaggggatttctttccagaagagaggacagtt-
tcttcaaat attgaattaaaagcaaaatagatgtaaaccgttgttggttttattgtt-
gaattccagGTAGACAAAAATCTTC GATTCATGATCTTGATGGATGGCGTTCATCCT-
GAGATGGACACTTGCAT TATTGTGGAATACAAAGgtattttcttgccctcatcagca-
tgaaattgctcttggtagaaaggataataata gttatccaaaacatcatcctatgtt-
catctgtttcttccctcttcattttccatagagtacagtatattctatctctgtcttaggaaa
atggactgtcattcatataatcttacagagaatcaattagtaatgtactctatgccgtgacaggtgcg-
aaggttttttttgaag gcaacagataaaaatatcctatatttcacctattgtaattt-
ccttaaaactgacattattgaataaatgttttactttcatcttgaa
tattattatgttatggaatcatacactttaccccaataatcatcgaaaagaatttccaaaaggttgagagagt-
tgtgttgatct gattactttcctctgcatcctttgagcttaacctttgaatatagtt-
tgctaaggaaagtagtctgtttatgatcctggagtggaa
tcaggctaagtgtcctcattcagaacccactgaatcagacagaatgaatttatttccttgaaagttcaaaatg-
tgtcactcaa gagtataaattttcaaatcttactctctcttttccttggatgtgagc-
aattcttcgataattgaatgaggcagattatatagactt
acatggaagactgttggcctgagaattcaaactatggtgttcaagacttcacngngagtccgatgccatttgt-
ttcccaca gGTCATAAAATACTCAATACAGTGGATTGCACCAGACCCAATGGAGGA
AGGCTGCCTATGAAGGTTGCATTAATGATGAGTGATTTTGCTGGAGGAG
CTTCAGGCTTTCCAATGACTTTCAGTGGTGGAAAATTTACTGgtaattctttatat
caaaatgatgccaaggagttggcatggcactttgctaaatgctgtgtgaatcaatacaaagataat-
taggacatggttctt cctcacaagaggtgtgcaatcttattgggaaatcatacttgc-
aagtcacaaatatagactaaagtttccagctgagaatat
gctgatggagcatgaaacactaaggagacagggagaatctcaggaaaaatcaagaataatttggatcaaatgg-
attcct gacatagaacatagagctgatcagaaagagtctgacattggtaatccaggc-
ttaagtgctctttgtatgtggttcagaaca gagtgtgggcagcctgagggggataca-
tacccttgacctcgtggaaagctcatacgggggagggatgaggctaagg
aagcccctctaaagtgtgggattacgagaggttgggggggtggtagggaaaatagtggtcaaagagtataaac-
ttcca gttacaagatgaataaattctaggggtataataacagcatggcactatagat-
agcatattgtactatatactggaagtgctg agagtagatcttacatgttctaaccac-
acacacacacacacacacacacacaccacacacacacaccacacacacaca
cgtgcacacaaacagaaatggtaattatgtgaggtgatggcggtgttaactaactttattgtggtcatcattt-
agccatacat gcatgtcatgaaatcaccatgacaccttaaagttatgtaatactaga-
tgtcagttatatctcaaagctagaaaaaatgt ggggaccaaggcagaagctcttctg-
ctctgtgtctaagggtggttctggggctgggatggggaggatggttaagtggt
atatttttttcatacctttgctcagtactatcattgtaagtgttcaatatatgtctgcttaataaattaatgt-
ttttagtaagtaatctc tgtttagtaatgtgtcagaaatgccctacttgcaatagga-
agaaaacctgtccagtcccttccttttttctgtaagtctgatttc
attgcctcccagaatgcatcaccatgtgagagatagagggaaggtgctgtccttatggggttaacagtgtgac-
taggga ggcaaaatatacctactaaagggtggtagcataattcagttcttatgtgag-
tatgtgtatgtgtgtgagtatgtgcacatgca catacattttaaaaggtctgtaata-
tactaacatgttcatagtggttacacctagcttataggtaacattttttcccctgtatcctt
gtttgtgtttatcaaattttcataacagtaatggtagaaggagtacctgacatggtaccatacatgctn-
ggncctgcctaatt tctcnatttcctttattgcccatacccccattgcttgacaagc-
ataagtccatactggcttgttttcgttcctcagactcagtac
accatgtagctccatgccctgggtctttgtatgtgctatttctactgcttagagtgctattgcccctgaccac-
cacgtggtca gcaacttctcttctgcgtctgtgtctatggtctatgattccagatgt-
catcttcactaactacccttctaatatgcccttccatcc
cacccgtcctcatccttaccccagccactctctatttggtggctctgttttattttcttcctagctcatcact-
ctttgaaatgaac ttatttacttattcaatatttgcttctttcactagaatgaatgc-
tccatgagagcagggacctgctttatcttgctcgccactgta
ttcacagtgcctagaactacgtctggcacatagtaggtgctcaataaatatcgatcaaatgaaagaatgagca-
aacgaac aaatgaacaacacgtgaggtaggcatcatgattccatcaacagaggagaa-
aaccagacttaaagnaatgaagtggng gagctgcatttgatcttgactgactccaac-
atccatgctcttgaccactgtgcatctccagagtgtaatgaacatactttactt
ttatattccaccaaaataacaaagccatgcccatgttagtagagagttaatcgacagtgcccttaaaatatgc-
atgcaccc agggtacaactatgcatgctgccctgtgttttcagttggatccaaatga-
attgccgtaaacaaagaggggattcaatgtctt tgactagtttgggatattttccta-
gtaaccaactttgcaaaataaagccactaatgacaaggagctttgttctacttctgcatc
actcaactgtcaatttttatctcttgcaagacttctaatctactagaacttttgtttttctgtgatttctg-
aacagagaagactaat ccaaaccctgtcattccagAGGAATGGAAAGCCCAATTCAT-
TAAAACAGAAAGGA AGAAACTCCTGAACTACAAGGCTCGGCTGGTGAAGGACCTACA- ACCCA
GAATTTACTGCCCCTTTCCTGGGTATTTCGTGGAATCCCACCCAGCAGA
CAAgtatggctggatattttatataacgtgtttacgcataagttaatatatgctgaatga-
gtgatttagctgtgaaacaaca tgaaatgagaaagaatgattagtaggggtctggag-
cttattttaacaagcagcctgaaaacagagagtatgaataaaaa
aaattaaatacaagagtgtgctattaccaattatgtataatagtcttgtacatctaacttcaattccaatcac-
tatatgcttatac taaaaaacgaagtatagagtcaaccttctttgactaacagctct-
tccctagtcagggacattagctcaagtatagtctttattt
ttcctggggtaagaaaagaaggattgggaagtaggaatgcaaagaaataaaaaataattctgtcattgttcaa-
ataagaa tgtcatctgaaaataaactgccttacatgggaatgctcttatttgtcagG-
TATATTAAGGAAACAAACA TCAAAAATGACCCAAATGAACTCAACAATCTTATCAAG-
AAGAATTCTG AGGTGGTAACCTGGACCCCAAGACCTGGAGCCACTCTTGATCTGGGT- AG
GATGCTAAAGGACCCAACAGACAGgtttgacttgaatatttacagggaacaaaaa-
tgatttctgaa ttttttcatgtttatgagaaaataaagggcatacctatggcctctt-
ggcaggtccctgtttgtaggaatattaagtttttcttgac
tagcatcctgagcttgtcatgcattaagatctacacaccaccctttaaagtgggagtcttactgtataaaata-
aactattaaa taagtatctttcaactctggggtggggggggagactgagttttttca-
cagtcctatataataattttcttatcctataaaataat
taggagttcccgtagtggctcagcaatagcaaacccgactagtatcgatgaggatgcgggttcgattcctggc-
ccccct cagtgggttaaggatctggcattgccgtgagctgtggtgtaggtggcagac-
acggctcagatcccacgttactgtggct gtggcataggccagcagctccagctctga-
ttagacccttagcctgggaacttccatatgctgtgggtgtggccttgaaaa
aaaataaataaataagataattactcaaatgttttccttgtctcagaaccttacttcaggataaagagtgaga-
aagtttttttta tgaagggccattattacagctcaaaaataagttgtcttcagcaag-
tagaaagcaataagcctgagagttagtgttcctatc
agtgtaaatattacctcctcgccaatccccagacagtccatttgaacaattaacggtgccctgggagtacagt-
tcagaaa cattaatgtggatgttccagacctgtatttttataagtacttgtcttgag-
ccggatggaaccatcattcctcaccattatttaga
agtggactgtgactctgttggagatcagggcacacggttaccaaaagcacacccttctcctggccttaccttt-
gcaaagc tggggtctgggacacagtcagctgattatacccttttactaacttcccac-
agctcaaatctggtcaattctccttcacaaatc tcttaaaaatccatcactcacctc-
cagcctcttctgctgtggccttgattcagcctctcacaatttttttttaaccagaattctgg
cagtggcccctgacttgcctctgtgctcccagccccgctgtcctctgatccatcctccatgccagcctt-
tttcaatctgctg gtcacgattcattgatgggttaggaaatcaatggcatcacaact-
agcatttagaaaaaggaaataggcgftcccgccgtg
gcacagcagaaataaatccgactaggaaccataaggttgcgggttcaacccctggccttgttcagtgggttaa-
ggatcc ggcattgccgtgggctgttttgtaagtcacagacatggctctgatccggca-
ttgctgtggctctggcgtaggcctgcagc atcagctccaattagacccctatcctgg-
gagcctccatatgctgcaagtgcagccctaaaaaaaataaaaaaataaaaa
aaaataaataaagaagtagacaaattgtatagaacaaccctgagtatgttgcctgagcacatataacaagggt-
aagtat tatttcaggaaactctggtttcacagatactcttggcatatggacccctag-
agtcctgatgtaaaatatattcttcctgggatc ttaggcaagaagt-ttgaaagctc-
caactctgcactgctgccaaagaaatgatttttaagtgcaaaactcttcccgttccctt
ccctgtataaaattccataggatctctccagtgcctctaggataaaggcagttttcattctctagttcaagg-
tgagagaaga ttttaattatttcacgttttagtggggaattcaagagtctggcacct-
gacatttgctgaactctctccattatccctctctagttc
cccagacgcatcctatggtagaaattcgcaaactagagtgagcgtcagagtaacccaaggaaactgggtaaat-
gcagc tccctgggctctaccccctgagattctgattcagtagatctgaagcagagcc-
ctggaatatgcatatgcatcattgtgtcac accaagcattctgggtaatgagagttg-
atgttaggttctcagtagtaagacaagtatagagattccgggggactgagtgc
tcagctctgccttggggaggagggagagggctaaagagaacaggagatggggacagggaatgctcaacctcca-
atc ttaggcatttgagctatgtcttaggggtcaggaggaggttaccaatatagtgat-
taagagattgaggttccagtcagagg gatatgctggagaaggggggtgaaaataatg-
tcataggtttggtgagtgcagatactttgagttttttaatatttttattgaaa
tatagttgatttacaatgctcttagtgagtacaattactttgaataagtgcatagatgtatgccattcttcca-
gaaatgatttatt gagctcctttgggcatcatgctaagtacaggggaaacagctgtg-
aagaggtccttcccttatgaagtcattcatccccttc
agtaaatgaaggtaaaggaaaaggatgagacagggacgccgtgttggaccagggtcagaaaggccttataaga-
cctt gcctggagggcaaggaacttgcctgtgagtaaggagagcttgagaaagcgata-
aagcaaagaaggaacattactgca ttgtgttttagaaaaaccatgtcctggggaaga-
actcctagagtcaggggggccagttgggagactgtgcttttttccagg
aggagataagtgaggctgctggctgagatggagcaaggatttagagaagcagatatgagattcatttagaagt-
tagaca ttttaggatctgacacataatttatcaccaaaaccagtgcatctctggctt-
tgggccaccagttttggagaagtggaatgta gggacctaccattacctgccaatctt-
tactacacagatgcctatttccctcctcatatttcctttctccagatcacgtcctattc
tattgccaggactcaagattccaccttgcatgcagtgatccatcttcacactggatggacagctctagggat-
gtcagagc acactcttgtccatactgctgactgggtctcctgtcagcccatctgtct-
atcagctgtggtattattagtataataagagggc tgtatatgagagacacaaaattc-
tagtgtagctcaaagataggctagagttattcctatgtacaacaaatatttatgggac
cccttctgtgtactgtcatggttgctgctttcatcatacttgtagtctaatggaggtgggggcagggcaggaa-
taagcgga tgtccacaaaatcagtaagaccacttatattcaacattttcataattta-
gttatttgagcccaaagggtccacatccgtggtat
tccaacttttttttccccggacatggatctttatctttttttttttttcttttttgcggccagacctgcggca-
tatggaagttcccag gccaggggttgaatgggagttgcagctgcctggtctacacca-
cagccacagcaaggtgggatctgagctgcatctgtg
acatacaccgcagctgaggtaacaccagattctgaacccactgaatgaggccagggatggaacccgtctcctt-
atgaa cactatgtcatgttcttcaccctctgagccacaacgggaactccagacttcg-
tctttaaatgtattctgacttggagagctat cacactaagcaattaacaggagctga-
cctggtttaggctggggtggggccctactcctcaatgttccctgaggcacatct
gtgggacccctgggcatcatctatctgagcagccttagagctgctcatccagttgactgttgatgtagaagtg-
caaacttc tgccttccttattgttgctttcttttttcattgttctctcccctttgtg-
tctttaagCAAGGGCATCGTAGAGCC TCCAGAAGGGACTAAGATTTACAAGGATTCC-
TGGGATTTTGGCCCATAT TTGAATATCTTGAATGCTGCTATAGGAGATGAAATATTT-
CGTCACTCAT CCTGGATAAAAGAATACTTCACTTGGGCTGGATTTAAGGATTATAAC- CT
GGTGGTCAGGgtatgctatgaagttattatttgtttttgttttcttgtattacag-
agctatatgaaaacctcttagtattc cagttggtttctcaataagcattcattgagc-
cttactgactgtcagacggagggcgtattggactatgtgctgaaacaatcc
tttgttgaaaatgtagggaatgttgaaaatgtagggaatgaaatgtagatccagctctgtttctcttttggag-
gattctttttcc tccatcaccgtgtcttggttcttgtttgttttgggtttttgtggg-
tgttgtattgtgttgtgttggttatggcagtgacagctattta
aactgtgaaacgggggagttcccgtcgtggcgcagtggttaacgaatccgactgggaaccatgaggttgcggg-
ttcg gtccctgcccttgctcagtgggttaacgatccggcgttgccgtgagctgtggt-
gtaggttgcagacacggctcggatcc cgcgttgctgtggctctagcgtaggccagcg-
gctacagctccgattggacccctagcctgggaacctccatatgccgc
aggagcggcccaaagaaatagcaaaaagacaaaataaataaataaataaataagtaagtaaaataaactgtga-
aacgg ggagttcccttcatggctcagcagttaacaaacccagctaggatccatgagg-
atgtaggttcgatccctggccttgctca gtgggttaagaatccagcgttgctgtgag-
ctgtgatgtaggtcgcagatgcagcccagatcctgcattgctgtggctgtg
gcgtaggctggcagctgaagctccgattcaacccctagcctgggaacatccatatgctgcaggtgtggcctta-
agagg caaaaaaataaaaaaataaaaaataaataaattgtgggacagacaggtggct-
ccactgcagagctggtgtcctgtagca gcctggaagcaggtaaggtaaggactgcag-
ctgggtaaggactgaattgcaccaactgggaagtaagcctagatcta
gaacttaagttagccctgacatagacacacagagctcaccagctaagtggttcagcttataagctggtcactg-
aaactga ggatgtccacaaaagcaaaataagtagcaacaggcagcgggatgcaagag-
aaagaggaggcctaaaatggtctgg gaatccctgccatacctatattttatcctact-
tatatttagtgcctgaatgtgtgcctggagagcaaagtttagggaaagcat
cgggaaatgcacagtattcatacccttaggaacaaagatcagttacctccagggtaaagactatttccaagtt-
taaatttca acccctgaacattagtactgggtaccaggcaacacttgccatcctcaa-
aatcaatgaatcctaaaattcaacctgggggt cagtgacagtctgtgacaaagtttt-
tgctggtcagtaacgaaataagtatgagcaccatctgagtatggtcaccaagatgt
caactctctttcctttggacgaattgtcattattccaagattaggtcctttctatttttgaggtgtgaaaaca-
tctttcctttcataa aataaaaggatagtaggtggaagaattttttttgttttttgg-
tctttttgctatttctttgggccgcttctgcagcatatggaggtt
cccaggccaggggtcgaatcggagctttagccaccggcccacgccagagccacagcaacacgggatccaagcc-
gc atctgcagcctacaccacagctcacggcaatgccggatcgttaacccactgagca-
agggcagggaccgaacccgca acctcatggttcctagtcggattcgttaaccactgc-
gccacgacgggaactcctaatgatactcttttatatttagctactat
gtgatgatgagaaacagtccacattttattattttttagccaatttgatatctcattactaagataatgataa-
ttttctctataaatt ttatttaagttagtgttatgaagtggttttgctagtgtagaa-
ggctaggatttgaattcagttcaagaaagaagagagggag
ggagggagagggatgggtagagggatggggcagtgggagagagcaaagaggagagacagtttttgtattaatt-
ctg cttcattgctatcatttaagggcacttgggtcttgcacattctagaattttcta-
aggaccttgaccgccagattgatatgcttct tccctttaccatgttgtcatttgaac-
agATGATTGAGACAGATGAGGACTTCAGCCCTTTG
CCTGGAGGATATGACTATTTGGTTGACTTTCTGGATTTATCCTTTCCAAA
AGAAAGACCAAGCCGGGAACATCCATATGAGGAAgtaagcaggaataccagtggaa
gtgcccctttcttccttccttcctaaataaacttttttattttggaacaactttagagttacagaaaagttgc-
aaagatattatag acagtagtgtttatatatatatataaatttttttttgcttttta-
tgaccacacctgtggcatatggaggttcccagtctaggggtt
gaattggagctacagctgccagtctgtgccataaccacagcaatgcaggatctgggccacgtctgtgacctac-
accaa agctcacagctggattcttaacccactgagcaaggccagggattgaacctgc-
atcctcgtggttcctagttggattcgttt ccgctttgccgcaatgggaactccaaat-
tattgttaatatcttactttactggggtacatttgttacaaccaatactctgatact
gaaacattactgttaactccgtacttgcttctttttgagtcatttgcaaagactggcttcttgacctgcttcc-
ttccaaacagct ggcctgcctatgctgttctcagacctgcaagcactgatctctgcc-
ccccttgccttctctccagtggtgtctccttccccaa
acaaacccagtgtggctctggaaagggagttaagtcaacataaaccaacacatattttgttgagctccaattt-
tgagcaaa tccctcacctacggcagacaggcatgatgttaagaactagggctttgga-
cacaaggtcaagaccaagaagggttcctc acccctactgattcagataaccaataat-
gaggctttgaatccctgtccaaaggttgttttttttcccttctattgagcttcttgcc
accttatcagttttttttatgacagtcaaatgacatgatatatgtgagcatacatggtaatttttaattctat-
ataaatgaatcact aaataaattaggaggatatatagtccacctttaagcgtattac-
acgtgtcacatgaatgtggcgacttaattgtagaggtt
taaatgtagcttcctataatagatgtgttcctaaactacattttaatcattggacttgtatttttatgttagc-
acttgctgttgaag aaaagcctatgccaaaagttcagtgaaaccaataatccactgc-
cagctttctgagttaaaaaaaatccctgggttttcaca
cacaggaacaccctgtgtgaaacactcatttagagcaaaatgcatctgataaggagttcctgttgtgcctcaa-
ctggttaa ggacctgacattctccatgagaatgtgagtttgatccccggccccactc-
gatgggttaaggatctggtgttgccacaaac tgcagctccgattcatctcctagcct-
agaaacttccacagcccagaatatgccacagaattcggctgtttaaaaaaaaaa
agaaaaaaaaaagaatcataaatgtgttggtttgttcaccaaatacatgataacttgctcttgccaagctcag-
cttcataaat attaagtcatttaatacagcagccaccttatgaacagatattactat-
acttcccatttacagataaggaaaatgccatatttaa
ccaagagattaaataactttcccgaggtcttatagcaagtaaatcatggtgcaggggtttgaccacacgcagt-
ctatctcc agagtctgtgtatttagccactgttttactttcaaatttaaatttataa-
aacttctaaattatctgttaaccataatctttggaatttt
taaaaccacgagttcctataaaatgtttcattgaaagtaagtcacttttccatagcttttgataatacatctg-
taggataaagta agccacagctctcttgcagacttggtacaccctggggcaaagcat-
catgcctgtcacgtacatggtggtccttactttgac
tctcagtgcttttattgcccaggaattttgtgagatttctagttgttgaggtttgtttaaagaggttatgccg-
gtacttggaaga gctcttttcttgctacctggagccttctcatatttcctttttgag-
gagggacatgaattgcctttcaaactcataaatatattttct
agtacacaagtctccatcttccttagacgcatggctcctggagttctccatcctcctgctccactttgggtgg-
gctcctctct gggtctgccaccaatctgccacccagagacatccttgacccacttcc-
agaccccaccatggcttcactttcttcgctttcct
cctttgtggaaccttctgcttaagaatctgaggaagaaaatttgcacgtgagctaaactggaggtactttcct-
gcctggtct tgcacgatagcttggctgagcccatgatgctgggtggctgttactttc-
catggacacccgaaggcgttgctcctttggctt ctagttgcatgcaggttgcttatc-
ccaggctgatctttcttccactgtaggtgacttttaagaattaagggattaatctatatc
tacaacaacaacaacaaagaccttttcaagctgaggtagggctttctgtatatgtttggagtggttatcca-
gcagactttac ttgaaggcaggggtcatatcctcaagtgctcataaacggaccacag-
aaagatctcataattgggtggagctgggtggg gaccgtgtcatgtggccaggaaatg-
ccagatgggaagggagtggcccttactgagctccagctgaactctgaattttct
agaaaactcagaaatctggatttttcatgtgtaatacccagatttatagatgtggaaagctaattcttttttt-
ttttaagggacta taggcaatgaactaagatctaggttgtatttggacaaggggtca-
tcagtttaagctgtgtagttgagcgctcagctattgg
gctgagggacccctaaatactgagacggggaggtccttgctctggggcatcacaagtacactccctggtctca-
ttcaaa cacttttcctacaaaattgatcccatttcttcagtgcactgtctgaatgca-
tttggcccagagccgtgctgaggcataggga aggggtccacggtttcatggcatcgt-
tttgtgctgtgtgtccctgctgtcgtccaggatacctacctctcctcctcctgcatc
tgaatgtccccccacagactctctgggattctacagcctctggcctgttcctcagacacctcttacctgccag-
ctttccaga ttcacattagttagtccaaatctactgccgtcagtgactcacttcatt-
tcttcttctccgaggcagttcagcccggtacagttg
ttttgtcaacacttcagttgagtctggaagatgtgcatgggttatgcacgagagcggtccatcattttgagct-
agaagtcctt tctcagcccagagacaagtcctcatctcctttacttcctgactcttc-
ttcctctgcatccttccaagatatctctttctccagcc
accacctaaatctcttcttttcccggggttccgtgctcaacccactcttcttcttaaatctgtggctgggtga-
acgcatctgct ggcaccacttctctgctaaagactccaaaaatccataggtcctgcc-
cggcctttgcccacctctctccaacactgtccag ctttagatgtagagctaatcccc-
ccagagatatcattccctggatgtctaagtcctttggtatctcactttcagcgtgttcaaa
atcctcttacaactgttctttctccttttccatcttgattattggcaacatgccagcctttcccctaccc-
ccagcagtgagcca agctagaaacaagggcttaatcttcaatctttccttctccatc-
cctaaacctaatgagtctccaagcccttcccagtttacac
cctaaatgttgctcaaaacatcccctagttcttccacgtgctctcctctatattgaaaggtcaagaaaggcca-
tcttccctcc actgtgaggaaatagatcttgatactgcccctgagctgggcagtcct-
cgacctgacaaactgtgcagtgtttctaaatctc tactggcaaaatgagagtgcctt-
tgacctgtgttgcgatctcagatcacagtggatgtaattgttttataggaatggtgaac
gaaaaagaagtaaatccctaatgccaaactcctgatcattctatgtcatttaatagcctgtcatttatgata-
aagtttcctcta ctggcattagcacaatacttctcaggaaaaaaaaatatgatgcca-
gatactgaaaagctcctgggtaaacatgaacatgg
gtaccgataaaatggtgaagccagtccaatcttagagtgacttcccttcatgctacttcatgctctttttttt-
ttttttttttaaga aaaaccccttttttttttctcacaccagtcacagaggagaccg-
aggcttagcaaggttaaggtcacatgattagtaagtgct
gggctgaaactcaaaaccatctctgcttgtctcctaaccctgtgcacctctgactattcaacagATCCTGTGT-
CA GGAGTTGGGATTCTTTGAAGgtaagggccttgaccaccgaattaaggtaatcttg-
ctctgtggcagg ccttgttttcagtattttaagtacactggctcaggtaatcctcac-
aacagccccaggaggaatgttctattacctccactgtat
agatgaggaacttgaggcacagaatggttgccaaggtcacacagctatattgggggttcatacccagccatcc-
aactct gtctgtactctctgccactctgcacccccagctcctgatccacttcctgtt-
tccatccctcgatttctgctgcactcaggggc ccctctccccctcggcctgtgagat-
ctgcttcagtaggcttttctccctgactcctccatccctgtccttacaggcagctgct
tctctccgggacacgaggggtccatacggacactctctactggctgggttgcgcctaactcgtgattcctcct-
ctgtttca gATTCGGAGCCGGGTTGATGTCATCAGACACGTGGTAAAGAATGGTCTG
CTCTGGGATGACTTGTACATAGGATTCCAAACCCGGCTTCAGCGGGATC
CTGATATATACCATCATCTgtaagtccgaaaatgcctgtcgtgtgtgccttaggctgctgcggag-
gag gccagggctatataagcagagtcagtgactgactgtgccctgcagtgttgatgg-
ccatggagattccaccgttagagctt ttttctttgttaaccttgaaggcaaatctgg-
ttaggaagataactttcaaagagtcaccatctggacattcatgcccatgtgct
tcaatcctgtatacaagcagtttagagtacagggaagggaaggacattatgaaagggagagggtgtgtttgga-
tccagc agctccatcctcagaatttatctgaagacactgcaaaattactaagaatca-
ctatgacaagaatgaggatggggtgatatg gcaaagttgtgatcctggaagaccttc-
atctcccatgttgcccaactctgaacatgaatttggtgaactagttggttaaggg
gatgatcctccaagtttctccctggttgagctccaaaaaccatgtaagtttctcatagcaaaaccgtataggt-
ccttagggc tttagttggaatatttgtgctgaaatgctggaaagccccatttgccat-
ttttgtatttgcaaaataatcatcaagaggggaga
atgcattctttcatgaccactgaccctctgaaaaggtcaggaatttagtctgaagtaggcaagcctcctaccc-
cgcttctg ccatgagcttgcacgcacaggcctgtcttgacatttcttctttatagat-
ttctttttgaatatcttgaaattgctttaaaaatattta
aagaatgtagaattatataaaataaaaaggaaataaccccacacctcccacaaaaccctgtttcctgcctttc-
tccaccca ctctccagggtaacacttggtaacagcatagttgtatcaccccaggcct-
atttttgagcatatcagcatttcaagaaatgtat tttttctcaataaaacatccctt-
atagttgaggaggggaggttatcattcctgggttttgttttttttttttttttaatgtaatcctggt
acatcggtaatttgcattttttattcattaatatctttggtatttctagtgttgggacacacag-
gtcaacctcagtttttgggtttttt tttttgtctttttgtctttctagggccacacc-
tgcagcatatggacgttcccaagctaggagtctaatcagagctgtagccac
cagcctacgtcatagccatagcaacgtcagatccaagccgtgtctgtgacctacaagcacagctcatggcaac-
accgg atccaccactgaacgaggccaggggatcgaacacacatcctcatggatccta-
gtcatgttcattaaccactgagtca tgatgggaactccaacttcaactattttaatg-
tctgtaaaacattccatttggaaaccatttcatttgtaaagcaaaatgaaaa
cattttgttcattttcaacagagttcgtagctgacttctgttctggaaaaaaggaaatggagcaaatttgagt-
gagaaagatt caaagataacttttcttttaaaaaaaattatatcttggaaacttctg-
ggctattgattctgaagactatttttctatatactgttttg
atagcaaagttcataaatgtgaaaggatcctgcgatgaatcttgggaagcagtcatagcccaatatatctttg-
ttgcttttaa aatgagatttagtttactaaatatttttctgatcataaaaataacac-
agatctaccgcagaaaatttggaaaaaaaaaaacttt
taaattcaaaaaacagttaaaccacaaatgatcccaccatccagagagcaatttgtactttggtgtctagttc-
atctttcttttt ctgtttacaagcacatataccacaagcattttttcaaaaaatgaa-
aatgggataatactatacatacgtctgtacacctgcat
agttactgaacagtctttgatctaccctgtaagtttctaacttttcattatttgaaatgatgttttggcaaag-
aaatatgtaggtg tgtctcgcacactttcataatgatttcttaggataaatttctta-
ggataaattcataatgatttcttataataatccatactctgcc
aactgatcttcagggaagccaactcgccttctcagaaataacatataacccatttacttgccctctcaccaat-
actaggtcc taatgtttttgtgtacagattctatatttttacatacaagaattcctt-
aaagcaaggcatgtcacagaaaaatagaaggaaga
cacaattgtcatgtttaaggactgcattctgtaccaaaaatgctaagttaaatgaacatctgaaacagtacag-
aaacgctat ctttcagggaaagctgagtaccaggtactgaacagattttggcaaata-
cagcaggcatggatgtttccaaaacatgttttt ctactttatctcttacagGTTTTG-
GAATCATTTTCAAATAAAACTCCCCCTCACACC ACCTGACTGGAAGTCCTTCCTGAT-
GTGCTCTGGGTAGAGAGGACCTGAG CTGTCCCAGgtaaagcatcctgcaggtctggg-
agacactcttattctccagcccatcacactgtgtttggcatca
gaattaagcaggcactatgcctatcagaaaacctgacttttgggggaatgaaagaagctaacattacaagaat-
gtctgtg tttaaaaataagtcaataagggagttcccatcgtggctcagtggtaacga-
accctactagtatccattgaggacacaggtt caatatctggcctcactcagtcggct-
aaggatccagtgatgccgtgagctgcagtgtaggccacagacgtggctcagat
ctggtgctgctgtggctatggtgtaggccggccccctgtaactccaattcgacccctaggctgggaacctaaa-
aagacc ccaaaaaagtcgctttaatgaatagtgaatacatccagcccaaagtccaca-
gactctttggtctggttgtggcaaacatac agccagttaacaaacaagacaaaaatt-
atcctaggtggtcagtgggggttcagagctgaatcctgaacactggaagga
aaacagcaaccaaatccaaatactgtatggttttgcttatatgtagaatctaaattcaaagcaaatgagcaaa-
ccaattgaa acagttatggaagacaagcaggtggttgtcaggggggagataagggga-
ggcaggaaagacctgggcgagggagat taagaggtaccaactttcagttgcaaaaca-
aatgagtcaccagtatgaaatgtgcaatgtgggaaatacaggccataact
ttataatctcttttttttttttgtcttttttgccttttctaaggctgctcccgtggcatatggaggttcccag-
gctaggagtccaaa cagagctgtagctgccagcctacaccagagccacagcaacacg-
ggaaccttaacccgctgagcaaggccagggatc gaacccgagtcctcacagatgcca-
gtagggttcattaaccactgagccacgacaggaattccagggtctgttgtgttctt
aaaacacttccaggagagtgagtggtatgtcataagtaaacaataaatgttaaccacaacaagcttatgaaat-
aaacagg aaagccatatgacctacaatcagtcattgggagaatccacaaaaggttga-
gcagaggatcaattccagctcacactcca gttttagattctcccctgccttaaagca-
tcacagactacataatctgagctgaagaataaaaattaaaactcaccccagtgc
aaaacagaaatgaaaaagtattaaaacgaggttcatactgttgttcattagcaatatcttttattcacagGGG-
TGCCC AACAACATGAAAAAATCAAGAATTTATTGCTGCTACGTCAAAGCTTATA
CCAGAGATTATGCCTTATAGACATTAGCAATGGATAATTATATGTTGCA
CTTGTGAAATGTGCACATATCCTGTTTATGAATCACCACATAGCCAGAT
TATCAATATTTTACTTATTTCGTAAAAAATCCACAATTTTCCATAACAGA
ATCAACGTGTGCAATAGGAACAAGATTGCTATGGAAAACGAGGGTAAC
AGGAGGAGATATTAATCCAAGCATAGAAGAAATAGACAAATGAGGGG
CCATAAGGGGAATATAGGG
[0090]
12TABLE 11 Contiguous 5' Genomic Sequence of CMP-Neu5Ac Hydroxylase
gene ctgccagcctaagccacagccacagcaacgctgggtc Seq ID No. 47
tgagccatgtctgcagcctatgccagagctccccgca
gcgccggatgcttaacccactgagcaaggccagggat
tgaaccctcgtcctcatggatagcagttgagttgttt
ccacggaactcttaggggaactcctgattatttttta
tttaaatttatatttctctgactttttcgtgtgctca
tcagccactgactgtgtatctccattagtcatggttt
gttaactctgtcattcaaaccctcttcatccttgcta
cgcagataacatcattataataaaatcgtgcctgaag
accagtgacgcccccaagctaagttactgcttcccct
ggggggaaaaagaagcaccgcgcgggcgctgacacga
agtccgggcagaggaagacggggcagaggaagacggg
ggagcagtgggagcagcgggcagggcgcgggaagcac
tggggatgttccgcgttggcaggagggtgttgggcga
gctcccggtgatgcaggggggaggagccttttccgaa
gtagcgggacaagagccacgggaaggaactgttctga
gttcccagtCCCGACGTCCTGGCAGCGCCCAGGCACT
GTTATTGGTGCCTCCTGTGTCCACGCGCTTCCCGGCC
AGGCAGCCCTGGCGGATCCTATTTTCTGTTCCCCCGA
TTCTGGTACCTCTCCCTCCCGCCCTCGGTGCGCAGCC
GTCCTCCTGCAGTGCCTGCTCCTCCAGGGGCGAAACC
GATCAGGGATCAGGCCACCCGCCTCCTGAACATCCCT
CCTTAGTTCCCACAGgtgagaaggcttcgccgctgct
gccgctggcgccggcagcgccctccacgcacttcgta
gtgggcgcgcgccctcctgcattgtttctaaaagatt
tttttttatccgcttatgctatcagttactgaggaag
tatttacaaatctactattattttgaatttgcctttt
tctccttatagtttatcagtatctcttgagactgtta
ttggtgcctgcaaatttaaaatgattggggttttatg
aggaagtgaaccttttatctttatgaaacgcctaact
gaggcaatgttaattgcttaaaatactttctttatta
tcagtgtggccatgccagtgtcctcttggttagaatt tgcctgat
[0091]
13TABLE 12 Contiguous 3'Genomic Sequence of the Porcine CMP-Neu5Ac
Hydroxylase Gene ctgccaaagctgggagatgggggaaagtagagtgggt Seq ID No.
48 tattgaaactgaatatagagttcagcatctaaaagcg
aggtagtagaggaggaagctgtgtcaacggaaatact
gagctgggttcacatcctctttctccacacagTCTAA
TGCCTTGTGGAAGCAAATGAGCCACAGAAGCTGAAGG
AAAAACCACCATTCTTTCTTAATACCTGGAGAGAGGC
AACGACAGACTATGAGCAGgcaagtgagagggggctt
tagctgtcaGggaaggcggagataaacccttgatggg
taggatggccattgaaaaggaggggagaaatttgccc
cagcaggtagccaccaagcttggggacttggagggag
ggctttcaaacgtattttcataaaaaagacctgtgga
gctgtcaatgctcagggattctctcttaaaatctaac
agtattaatctgctaaaacatttgccttttcatagCA
TCGAACAAACGACGGAGATCCTGTTGTGCCTCTCACC
TGCCGAAGCTGCCAATCTCAAGGAAGGAATCAATTTT
GTTCGAAATAAGAGCACTGGCAAGGATTACATCTTAT
TTAAGAATAAGAGCCGCCTGAAGGCATGTAAGAACAT
GTGCAAGCACCAAGGAGGCCTCTTCATTAAAGACATT
GAGGATCTAAAATGGAAGgtactgagaatcctttgct
ttctccctggcgatcctttctcccaattaggtttggc
aggaaatgtgctcattgagaaattttaaatgatccaa
tcaacatgctatttcccccagcacatgcctaactttt
tcttaagctcctttacggcagctctctgattttgatt
tatgaccttgacttaatttcccatcctctctgaagaa
ctattgtttaaaatgtattcctagttgataaacagtg
aaacttctaaggcacatgtgtgtgtgtgtgtgtgtgt
gtgtgtgtttaccagcttttatattcaaagactcaag
cctcttttggatttcctttcctgctctctcagaagtg
tgtgtgtgaggtgagtgcttgtccaaacactgcccta
gaacagagagactttccctgatgaaaacccgaaaaat
ggcagagctctagctgcacctggcctcaacagcggct
cttctgatcatttcttggaagaacgagtgctggtacc
ccttttccccagccccttgattaaacctgcatatcgc
ttgcctccccatctcaggagcaattctaggagggagg
gtgggctttcttttcaggattgacaaagctacccagc
ttgcaaaccagggggatctggggggggggtttgcacc
tgatgctcccccactgataatgaatgagggattgacc
ccatcttttcaagctttgcttcagcctaacttgactc
tcgtagtgtttcagccgtttccatattaggcttgtct
tccaccgtgtcgtgtcgtcaatcttatttctcaggtc
atctgtgggacagtttagtgcgaatggactcagaggt
aactggtagctgtccaagagctccctgctctaactgt
atagaagatcaccacccaagtctggaatcttcttaca
ctggcccacagacttgcatcactgcatacttagcttc
agggcccagctcccaggttaagtgctgtcatacctgt
agcttgcttggctctgcagatagggttgctagattag
gcaaatagagggtgcccagtcaaatttgcatttcaga
taaacaacgaatatatttttagttagatatgtttcag
gcactgcatgggacatacttttggtaggcagcctact
ctggaagaacctcttggttgtttgctgacagactgct
tttgagtcccttgcatcttctgggtggtttcaagtta
gggagacctcagccataggttgttctgtcaccaagaa
gcttctgcaagcacgtgcaggccttgaggtcttccga
cttgtggcccggggactctgctttttctctgtccttt
tttctccttagtgggccatgtcctgtggtgttgtctt
agccagttgtttaagggagtgttgcagctttatgatt
aagagcatggtctttccttgcaaactgcttggtttag
aagcctggctccaccacttagcggctctgtgacctcg
gacacatttcttagcctttctgggcctcgctcttctt
cctcataaagtgaaaatgaaagtagacaagccttctc
tgtctggctactgagaggatggagtgatttcatacac
ataaagcacttaaaataatgtctggcatatgatacat
gctcaataaatgtcacttacatttgctattattatta
ctctgccatgatcttgtgtagcttaagaacagaggtc
tttacaggaattcaggctgttcttgaatctggcttgc
tcagcttaatatggtaattgctttgccacagactggt
cttcctctccttcacccaaagccttagggggtgaacg
atcccagtttcaacctattctgttggcaggctaacat
ggagatggcaccatcttagctctgctgcaggtgggga
gccagattcacccagctttgctcccagatacagctcc
ccaagcatttatatgctgaaactccatcccaagagca
gtctacatggtacactccccatccatctctccaaatt
tggctgcttctacttaggctctctgtgcagcaattca
cctgaaatatctcttccacgatacagtcaagggcagt
gaccatacctgttccaccttcccttcctcagccattt
ttcttctttgtacataatcaagatcaggaactctcat
aagctgtggtcctcattttgtcaatctaatttcacag
cctcttggcacatgaagctgtcctctctctcctttct
gcctactgcccatgagcagttgtgacactgccacatt
tctcctttaacgacccagcctgctgaatagctgcatt
tggaatgttttcaatttttgttaatttatttatttca
tctttttttttttttttttttttagggccgcacccat
gggatatggaggttcccaggctagggatccaatggga
gctgtagctgctggcctacaccacagccacagcaatg
cacaattcgagccaatctttgacctacaccagagctc
acggcaacactggattcttaacccactgattgaggcc
agggatcaaactctcgtcctcatagatacgagtcaga
ttcgttaacctctgagccatgatagttgttagttact
cattgatgagaaaggaagtgtcacaaaatatcctcca
taagtcgaagtttgaatatgttttctgccttgttact
agaaaagagcattaaaaattcttgattggaatgaagc
ttggaaaaaatcagcatagtttactgatatataagtg
aaaatagaccttgttagtttaaaccatctgatatttc
tggtggaagacatatttgtctgtaaaaaaaaaaaatc
ttgaacctgtttaaaaaaaaaacttgactggaaacac
taccaaaatatgggagttcctactgggacacagcaga
aatgaatctaactagtatccatgaggacacaggtttg
atgcctggcctcgctaagtgggttaaggatatggtgt
tgctgcagctccaattcaacccctatcctgggaaccc
ccatatgccaccctaaaaagcaaaaagaaaggtgctg
ccctaaaaagcaaaaagaaagaaagacagccagacag
actaccaaatatggagaggaaatggaacttttaggcc
ctatctccaactatcacatccctatcaccgtctggta
agaaatggaaaaaatattactaagcctcctttgttgc
tacaattaatctgattctcattctgaagcagtgttgc
cagagttaacaaataaaaatgcaaagctgggtagtta
aatttgaattacagataaacaaattttcagtatatgt
tcaatatcgtgtaagacgttttaaaataattttttat
ttatctgaaatttatatttttcctgtattttatctgg
caaccatgatcagaaatctttaaacaatcaggaagtc
ttttttcttagacaaatgaaaatttgagttgatctta
ggtttagtacactatactaggggccaagggttatagt
gtgactattaaatcacagataatctttattactacat
tatttccttatactggccccacttggatcttacccag
cttagcttttgtatgagagtcatccttaaagatgact
ttattctttaaaaaaaaaaaaacaaattttaagggct
gcacccatagcatatagaagttcctaggctagcggtc
aaattagagctgcagctgccagcctatgccacagcca
cagcaatgccagatctgagctgcatctgtgacctaca
ctgcagcttgcagcaatgctggatccttaacccattg
aacaatgccagggattgaacacacatcctcatggata
ctgctcaggttcctaacctgctgagccacagttggaa
ctccaaagcagactttattctgatggctctgctgatc
tctaacacgttattttgtgccatggtgtttatcttca
ctttactcaagtcagggaaacacgaagagtctcatac
aggataaacccaaggagaaatgtgcaaagtcacatac
aaatcaaactgacaaaaatcaaatacaaggaaaaaat
atcttcactttcaaaatcacctactgatgatgagttt
atattttccttggatatttgaatattagctatttttt
ttcctttcatgagttttgtgttcaaccaactacagtc
gtttactttgatcacagaataatgcatttaagcctta
aatagattaatatttattttcaccatttcataaacct
aagtacaatttccatccagGTCTGTTAAATGCACAAA
ACACAACTGGAAGTTAGATGTAAGCAGCATGAAGTAT
ATCAATCCTCCTGGAAGCTTCTGTCAAGACGAACTGG
gtaaataccatcaatactgatcaatgttttctgctgt
tactgtcattggggtccctcttgtcaacttgtttcca
atctcattagaagccttggatgcattctgattttaaa
ctgaggtattttaaaagtaaccatcactgaaaattct
aggcaagttttctctaaaaaatcccttcattcattca
tttgttcagtaagtatttgatgagaccttaccatgtg
taaacattgcactaggtattaagaaatacaaagatgg
ataagatagagtcggcgtaaatgagatgatataatga
gacgttataatgaaactcacaattccagttgggaaat
aaagtccttcaaattccatgactctttctggcacacg
ttagaggctacagcttctgtgtgattctcatgctggc
tccacttccactttttccttcttccatactcaagaaa
gcctatagaaatatgagtaagaagggcttaatcatag
gaataaatttgtctctgttctaagtgattaaaaaatg
tctttatcagtataaaaagttacttgggaagattctt
aaaactgcttttacacactgttctagaatgactgtta
tataaataaaaaagtagatttgatctaacacaattaa
atgacctttggaaatattgactattctcaccttgccc
ctcaaagggatgcctgaaccatttccttcttttgcca
gaaagcccccaccctttgtctgttgacctagcctagg
aaatcttcagatcacgttgttagcacgaactggttac
atgtgctgtacaaatactatttattcatctgattaaa
aaaaaagagataagaagcaaaagtttgactatcttaa
actgtttgcgtaggtgagaggacaattgaccatctac
tttatgagtatgtaacccagaaacttaaagctcctta
agggagctaagtcttttggataagacctatagtgaga
ccttttagcaaaatggttaagactgaatggactgaat
ggagctcactagcgtgggttcatatcctgatgctcaa
acacgcaattaaatgactttaggtgggttagtctctg
ttccttagtttcctcaatgggagataatattggtagt
agcgattttactgggttgttgaaagaacatctgttaa
atgttcagaacgtgttacgacagagtacagagtaatg
atttgcttgtatatgtatgactcaaatagtctgccat
atgccttgtgactgggtcctgtggagcaggaaggagg
gatttcccacccagcagaaagttgggtaaactggaaa
atagactgaggccaggaaatgatgcaaagcgttgatg
ttcactgccacggcaggtgaagggcagggccagagtt
gtcagtagggtcaggggaggactggaaataaccaaga
cccactgcacttttcagcctttgctccagtaaggtaa
tgttgtgagagtagaaaattttgttaacagaacccac
ttttcagtacagtgctaccaatactgtagtgatttca
taccacatcccaagaaagaaaaagatggctcaatccc
atgtgagctgagattatttggttttattgttaaataa
atagcattgtgtggtcatcattaaaaaaggtagatgt
taggaaagtagaaggaagaagactctcacctacattt
tcatcactgtttggtatctgccagttgtcaccttggt
ccccttccccgcctctcccctgcctcctcttcctcct
tctcctttttttggaatacaattcaggtaccataaaa
tttacccttttagagtgttgactcaatggtttttagt
attttcacatgttgtgctattactatcactatataat
tccaggtcattcacatcaccccccaaagaaaccttct
aactattagcagtccattcccttcttccctcagcccc
tggcaaccactaatctacttactgtctccatggatgt
tcctatattgaatcaagctagcataaaccccacttgc
tcatggtcataattcttttttatagtgctaaattaca
tttgctaatattcaattaaggatttctatgtccatat
tcataaggaatattggtgtgtagttttctctttgtgt
gatatctttgtctggttgggggatcagagtaataatt
actgctctcatagaatgaattgagaagtgttccctcc
ttttctatttattggaagagtttgtgaagtatattgg
tattgattcttctttaaacatttggtcagattcacca
gtgaagccatctgggccatggctaatctttgtgaaaa
gttttttgattactaattaaatctctttaatttgtta
tgggtctgtcctcagacgttctagttcttcttgagtc
agttttgttcatttgtttcttcctaggactttctccc
tttcatttggattatttagattgatagtaatatcccc
cttttaattcctggctgtagtaatttgggtcttttct
cttttttcttggtcagtttagctaaaggtttgtaatt
gtattaatcttttcaaataactaacttttttgttttg
tttgtttttttgtttttttgtttttttgtttttttgt
tttttttttgctttttaaggctgcacctgaggcatat
ggaagttctcaggctagaggtctaatcggagctacag
ctgctggcctataccacaaccatagcaatgccagatt
caagctgcatctgcgacctacaccacaactcggccag
ggatcacacccgcaacctcatggttcctagtcggatt
tgttaaccactggtgccacgacgggaactcccgccca
tttttttttaacacctcatactttaacataaagatgg
gcttcacatggactgatagctcaaatgaggaaggtaa
gactatgaaagtaatggaagaaatgtagactattttt
gtgacctagagattactgataacttcttgacttttca
aacaatacttcaaaagtacagcccaaagggaaaaaag
aaagaaaaaagaacacacatacacaaacctagtgaat
aagatatcatcgatacactacagatttctatgaactg
gaagaccccatggacaaagttaaagaacatatgatag
tttgagtgattattttgcaatatttacaaccaatgag
ggaatattatccagcttataggaggaagtaatgcaaa
tcgacaagaaaaagataggaaacccaatataaaaatt
aagaaaatacaaaaattaagaaaggatatgaactagc
attttacaaaagaaaaatctccaaaagtcaatcagca
catgaaaatatgctcaaacctattaattattagaaaa
ctacagactgaagcaatgaggtgctttactttacatc
tttttgactgataaaaagttagaaacaaggtgatatc
aaatgtcagggataaaaggatatagaaatcgtcatgc
ctgtggtgggagtatggccggtgcagtcatgtgggaa
ggtaatctgacagtggttaggcagagcaggtttatga
atacactgtggcccatcaatcccacgcctgtttatgt
accaaagaaatcctgttgtggcagaatctatgggtcc
acccctgggagcatgaattaataaaatgtggcaccag
ggtgtgtgaaactccagctagagatgagatgtccaca
tggcaacatgaatgcatcttagaaacatagatttgag
tgaaaaagagtaagaaacagccgggaaacccaatacc
atttataaaaattaaagatgcacacatacaatgtagt
aaatattttgcatgaactttcaaatggttgcctacag
ggggggagagtaaagaagagtagaaaacaaagataaa
gggagtaagtaagtagctctgcctggactgaatataa
tgtgtcatgaactgagaaatatggttaacataatcct
cttaacttgaggtcctaaatgaatgaatgagtccact
attcatttacccattctttaatgtgtattgcattata
atccatttttttagaaccaacgaattttgttcccata
actactaatcagcctgccttttctccctcattccctt
atcagctcaggggcattcctagtttttcaaacgttcc
tcatttgaaccaaaaatagcatcattgtttaaattat
acttgttttcaaatacgatgcttatatattccaagtg
tgtttgcccattttcttaggtggtagaaatttttcat
tctacttttctatctactcagttttcccgttggaatt
attttccattgctattaaaacttagaagtcccccctg
tgatatgccatttttttcatactttttaagcacttgg
ttgcttttctttgtgtctttaagcacctagaatactt
ataaccattgcacagcactgtgtatcaggcagccctt
cctcttccactaatttatggtccttctcttagatata
ttaaactgttatttaattaggatcctctcttcgtcct
tagatttaattattatagttttctaatatgtttttat
tataattcctcttcattattcctccctattaaaaatt
ttaatgaattccatttgtttgttcttctagttaaata
ttaagtcataatccaaataacttagatgtcattagtt
tatgtggtcaaagtaaggataccacatctttatagat
gcaggcagttggcagatgtcatgattttcttcagtgc
ataaatgcaatttatctttgagcaaggggcataaaaa
cttttatggtattggctttgaaataatagttaagaac
tgcagactcagtttttcctgcttttcttgaaaaagaa
cacttctaaagaaggaaaatccttaagcatggatatc
gatgtaattttctgaaagtctcctgtaattccttggg
atttttgttgttgtttgttggtcggtttttttgggtt
tttgtttgtttgttttgttttgttttgttttgctttt
agggctgcacctgtggcatatggaagttcccaggcta
ggggtccaactggagctacagctgccagcctactcca
cagccacagcaacatgggatcctagctgcatctgtga
cctaaccacagctcttggtaatgccagattgttaacc
cactgagcaatgccagagcatcgaatctgcctcctca
tggacactagtcagattagtttctgctgagccacaat
gggaattcccaattccttgtatttttgaactggttat
gtgctagcatataattttgtttcttgatctttgtggg
ttttttttttttttgtctcttgtctttttaaggctgc
acccacagcatatggaggttcccaggctagaggtcaa
attggagctacagctgccagcctacacaacaactgca
gcaaagtggggcccaacttatatgacagttctggcaa
tgccggattcctaacccactgagcagggccagggatc
gaacctgagtttccagtcagtttcgttaaccactgag
ccatgatagtaactcctgtttgttcagtcttgaacct
cctttttaattctttattccttgagggtgaaataatt
gccataataatactatcatttattacatgccttctct
gtgctaggcatagtgacactttaggatttattataca
cttaatccctacaacaactctgcaaagtatgtatcat
aatcctatttgacagatcaggaaattgcagcccagga
tgcagataatatgcatccatcacaagtgactagcata
tagtccctctgctattcagcagggtctcattgccttt
ccattccaaatgcaatagtttgcatctattgtatatg
tgttttggggtttttttgtcttttttttttgtctttt
ctggggcctcacccttggcataggtaggttcccaggc
taggggtcaaattgaagctgcagctgccagcctacac
cacagccacagcaactcgggatctgagcctcatctgc
aacctacaccaaagctcacggcaacaccggatcctta
acccactgagtgaggccagagatcaaaccggcaacct
catggttcctagtcggattcattaaccactgagccac
gatgggaactccctaaatgcaatagtttgctctatta
accccaaactcccagtccatcccactccctcctcctc
cctcttggcaaccacaagtctgttctccatgtccatg
attttcttttctgggggaaagtttcatttgtgccatt
tttcattttacggtaatttttacttcagtttcttcca
ctagcagttgtcttaaagtgagtataattaatattca
tttggaaaatgtaagcaaaacattttttaaagggcca
tgcccacagcatatgaaagtttctgggccaggggttg
aatccaggctccaagttgcagctgtgccctacactgc
agctgggcaatgctggatcctttaacccactgtgccc
ggctagggatcaaacctgcatttccacagctacccga
gccattgcagttggattcttaacccactgcactacag
tgggaactcccacaaaacattttttaatgtcctttga
ataaagtaggaaagtgctcgtctttgagggcagggcg
gcaatgccatttccacaaggtttgctttggcttggga
cctcatctgctgtcatttagtaatgaataaaattgct
gacagtaataggattaactgtgtgtggagatagccag
ggttagagataaaaacactggagaagtcaaataagtt
gctcgaggtcctctagctaataagctattaagtggga
gagtgagggctagaaacaggccatctgtctcccaagc
acatgtccattagtggtttgctgatagccttccagaa
caacagagaggactctcaaacatggtcttgcctccct
ccaattgatcccctccatgtgcctcacagcgggtctt
tctaaaattaagttctgattttaattctcccttgcta
tagcacttaggtatggctttcagccgtgcaataaaaa
gcaggcaagagtggctcaatcatataggaggttgttt
ttcttagatcccaagcaggtaatcctgggcattatgg
ttgttctgcgtttatcaaggagccaaattctctatca
cctcctgttctatcctcctcagtatctggctctattc
ttcagcatctcaagatggcttgtgctcctccaagcat
ggcagtcaaattccacacaagagggggaaaatatgaa
gggcagacagtgctggtctcctgagctgtccctcttt
gtcggggaaataaagtattccttcaagtcccgtgaga
cttctgaagtagacgtctgcttacgtctcacccacca
gaactatgtaaactgcacatagtgctaggtctacata
gccatcataactgccagggggtgggaaatctttaaat
aggtgtaccaccacacaattaggatgctaatagtaag
ggagaaggagagaataggttttgcgcaagccaccagc
atgcctgccacaattgcttaaaattcttcattgaccc
ctcattgccacaggatgaaatccaaacgccttcttag
ttgggaatctgacctacctgtctctcccacctggttc
agacaccattctccttggtcataaaattccagtcatt
tgtgaacatccagctcccccatgcctccatgcctttg
cacatgctgttcttttatcttttatgttgtcctttta
tcttttatccaaaagagatatcccatcatcacatctc
ttttgtcagcccccaaatactttgtctttcaagttca
gctggaggattacctctcaatcattcatttgattgta
atgaagttgttggtggtatgcctgtgtctcgacacat
ctgcgatctcatgagttccttaagtggaatgtgaata
gcgggatgaacagtattggtcttcagccctcatctct
gcagatgttgcttgacccaaatgagcgttgcctttta
ttttgattttgctttgatttgtctactccatgtactt
gagccatgcatttctgtcttagcgatgctttttaaaa
gtcattttttggttgattatccagatttgtccacctt
tgcttctagTTGTAGAAAAGGATGAAGAAAATGGAGT
TTTGCTTCTAGAACTAAATCCTCCTAACCCGTGGGAT
TCAGAACCCAGATCTCCTGAAGATTTGGCATTTGGGG
AAGTGCAGgtaaggaaatgttaaattgcaatattctt
aaaaacacaaataaagctaacatatcaatttatatat
atatatatatatatattttttttttttttttacatct
tatattaccttgagtattcttggaagtggctagttag
gacatataataaagttattctgaagtctttttttttc
tttttccatggtgagcagtggcttgatgtggatctca
gctcccagacgaggcactgaacctgagccgcagtggt
gaaagcaccaagttctagccactagaccaccagggaa
ctccctattctaaattcttgagcacattatttaggaa
cctcaggaacttggcaggattacaggaaatatatcta
gatttaaaaaaaaatcttttaacagaggtcccaaagg
agagtcatgcacagctatgggaggaagttcagaaact
gcccttgctaccagatcactgtcagataaaatggcca
gctacatgtttctgcacattgccctaagatctttaca
aacttttctgtgcatttttccacttttaaaagaaaat
ttcggggttcctgttgttgctcagtggttaacgaacc
caactagtatccatggggacaggggttcgagccctgg
ctcactcagtgggttaagaatctggcattgctgtggc
tgtggcgtaggctggcggctacagctcagattggacc
cctagcctgagaacctccatatgccgcaggtatggcc
taaaaaaaaaaaaaaaaaaaaaagagagagagagaat
ttcctccagaaaaaacactttggtagtttgggagaag
taaacaaccaaaaattaatttttctggagtattcggg
aagcttgtaaaaatgggctcttacttttttgaggaga
caaatgggaacctacccagaagaggcacaatcacctg
catttgatttcttgacctctccctaccttctttgctg
gctttccacatttggatttctgtgaccttatctctgc
tccttggtgttttcatttttcctgtggacgtgccaga
ctatgggaagggagtaaggcgttgatttagaatcctg
tagtctctgcctgtctctagtcattgttttcaccctt
ctcaaaggaccttgacatcctgagtgagtccgcaagt
aatttaggggagaagccttagaagccagtgcagccag
gctacatgactgtgtccacccactggaaccagtcatt
tttatacctattcagcccccctaccatttaaatcccc
agaggtctgccataacatctgtaactccctttcctgg
taaattgtgttctaaaagactggtaacaaaagatatt
ctgtggtacagagcataattaaatacctgggagctga
tttgagtggggtaaatcaactggtttgacccctaaaa
cccaccatgagcatttctgttctaataaagtaatgcc
cgtgctgggaattgtgttctacggaaatgctcctgct
gtgtctttcttgagtcctgtgtcattgatcatctgaa
tctcactgtcttccaacagATCACGTACCTTACTCAC
GCCTGCATGGACCTCAAGCTGGTGGGACAAGAGAATG
GTGTTCGACCCTTGGTTAATCGGTCCTGCTTTTGCGC
GAGGATGGTGGTTACTACACGAGCCTCCATCTGATTG
GCTGGAGAGGCTGAGCCGCGCAGACTTAATTTACATC
AGTCACATGCACTCAGACCACCTGAGgtaaggaaggg
tgagccctcaatccgaagaaaatgctgcaataaaagc
actgttggttttcagctttttttgtaatcactgctca
ttctgaggtagattcgcttgggctgataaaaagagaa
ctattcagataaatgcttgcatttgcatagcctcttt
tttaaaaacttttttttttttttggcttttcagggct
gaacctgtggcatatggaggttcccaggctaggggtc
gaatcagagctgtagccccgggcctatgccactgcca
tagcaacatgcatagcctcctttttaaagtgccttcc
tgttttataccattgggatgtgagaagagctattgtg
gaaangaagcatggtnataaccctggacctctcacgt
cctaccctcaggntagtgggaaaactctgagtttaag
gacatcaaagtgactcctttttagttacattatggng
gaatcagcncatatttttacaaggggcggagngtaan
ctgttggagtttacaagacatatggtggcattgcaac
tacttaaccctactattatagcacaaaagcagccata
gtcggtcctgaaggagcctgatgccttcagctttata
ggcaatgacgtgtgaatatcacaaacagtttcctgtg
tcaccaaacatgattgccttttgatttccctttcaac
cctttaaaaaaaaggtaaaagcccttcttagcattca
gcagcaggtcgctgtgttttgccaactcctgatctgt
agcatttcgacaacactgagctctcaacttttgaacc
tgagtccaccacatccttcagtgaaaccagagccatg
tgatactaaggatagaacggaaacttcctgaatccag
gcgatcaaataggaggggagaaagaggaactttcatt
gacaaaaccacaaatattgtgaatggactgttacaaa
tattgtgaatgctcctattcccaaccccctggcttca
ttacagggtcctatgtgttcatccttattgagaaatt
tgtattgctactgccaggttgccaatacccagcggtg
cccatggtgttctaaaatgaagcaatttcaactttat
tttttttcctgtgactttacatgacaagttcacatga
aggatatactttgatagtaatgtccatggttagggaa
tatacattgtttgctggttgactggcccctggatttt
tctattgaaagtccatgagatctcgaaggcacaggtg
tgttctctcgctttttaaggaaagggtttaaaaactt
aagtaattaacagctttagtaacaaattacctataac
acacttaaaaaccgaataccacccactggagtattgt
gctacgattaaaaatctacttgtctactacatgatat
ctttgtcccacagaaggttctggaaccaaacttgtaa
tttcaggattatgagagccctgagttcacgcattgtg
taataactatgttgtgtgtgtagtcaatttgtacagc
ttgcttagagagaacaatgtcaagttaaggaggcgat
tgctttatagtgcctgtcacaagatgccattgccatt
gtcctagcaagagatattctatgggagtatactacat
tttagtgaggataagaactttttatggcatttagtcc
ggtcatttcccaaccactgtcctgaaaaccaatttca
ttttgatttcaggggcttgtgtgggcaaagttgccag
gcattaaaaagccacttctcaactgtgagtatcacaa
tgctttagttggtgaccctgagttttttgactcttac
ctgctcaactaataaacacagagtcatttatctccat
tgctcttgtctgacctttatttaccgaataatgcctt
atgggttcacaaaacaagggggaggggccacatgcct
tagaactgtctttagtcaagaaatgngattttattat
gtaaatatatgagtattataatagatagtgttattaa
tagacaccagcaagaattgtcaataatttaaaaatca
caaattaaaatacatccatgttagnatcatttatcct
aactcccaaagccctttaaagtggaaagatttagatg
ttaacccagagattaaagacatgttcaaagaatcctt
gattttttttgaatcccttgtttttagagaagaaaac
ctaatgattttccccctctggattctacatattaaat
atagttttggaacttgaatattagtatggttaataag
tgctgatatgctgaattttgtttttatttttttcatt
ttccatagagtacagtatattctatctctgtcttagg
aaaatggactgtcattcatataatcttacagagaatc
aattagtaatgtactctatgccgtgacaggtgcgaag
gttttttttgaaggcaacagataaaaatatcctatat
ttcacctattgtaatttccttaaaactgacattattg
aataaatgttttactttcatcttgaatattattatgt
tatggaatcatacactttaccccaataatcatcgaaa
agaatttccaaaaggttgagagagttgtgttgatctg
attactttcctctgcatcctttgagcttaacctttga
atatagtttgctaaggaaagtagtctgtttatgatcc
tggagtggaatcaggctaagtgtcctcattcagaacc
cactgaatcagacagaatgaatttatttccttgaaag
ttcaaaatgtgtcactcaagagtataaattttcaaat
cttactctctcttttccttggatgtgagcaattcttc
gataattgaatgaggcagattatatagacttacatgg
aagactgttggcctgagaattcaaactatggtgttca
agacttcacngngagtccgatgccatttgtttcccac
agGTCATAAAATACTCAATACAGTGGATTGCACCAGA
CCCAATGGAGGAAGGCTGCCTATGAAGGTTGCATTAA
TGATGAGTGATTTTGCTGGAGGAGCTTCAGGCTTTCC
AATGACTTTCAGTGGTGGAAAATTTACTGgtaattct
ttatatcaaaatgatgccaaggagttggcatggcact
ttgctaaatgctgtgtgaatcaatacaaagataatta
ggacatggttcttcctcacaagaggtgtgcaatctta
ttgggaaatcatacttgcaagtcacaaatatagacta
aagtttccagctgagaatatgctgatggagcatgaaa
cactaaggagacagggagaatctcaggaaaaatcaag
aataatttggatcaaatggattcctgacatagaacat
agagctgatcagaaagagtctgacattggtaatccag
gcttaagtgctctttgtatgtggttcagaacagagtg
tgggcagcctgagggggatacatacccttgacctcgt
ggaaagctcatacgggggagggatgaggctaaggaag
cccctctaaagtgtgggattacgagaggttggggggg
tggtagggaaaatagtggtcaaagagtataaacttcc
agttacaagatgaataaattctaggggtataataaca
gcatggcactatagatagcatattgtactatatactg
gaagtgctgagagtagatcttacatgttctaaccaca
cacacacacacacacacacacacaccacacacacaca
ccacacacacacacgtgcacacaaacagaaatggtaa
ttatgtgaggtgatggcggtgttaactaactttattg
tggtcatcatttagccatacatgcatgtcatgaaatc
accatgttgtacaccttaaagttatgtaatactagat
gtcagttatatctcaaagctagaaaaaatgtggggac
caaggcagaagctcttctgctctgtgtctaagggtgg
ttctggggctgggatggggaggatggttaagtggtat
atttttttcatacctttgctcagtactatcattgtaa
gtgttcaatatatgtctgcttaataaattaatgtttt
tagtaagtaatctctgtttagtaatgtgtcagaaatg
ccctacttgcaataggaagaaaacctgtccagtccct
tccttttttctgtaagtctgatttcattgcctcccag
aatgcatcaccatgtgagagatagagggaaggtgctg
tccttatggggttaacagtgtgactagggaggcaaaa
tatacctactaaagggtggtagcataattcagttctt
atgtgagtatgtgtatgtgtgtgagtatgtgcacatg
cacatacattttaaaaggtctgtaatatactaacatg
ttcatagtggttacacctagcttataggtaacatttt
ttcccctgtatccttgtttgtgtttatcaaattttca
taacagtaatggtagaaggagtacctgacatggtacc
atacatgctnggncctgcctaatttctcnatttcctt
tattgcccatacccccattgcttgacaagcataagtc
catactggcttgttttcgttcctcagactcagtacac
catgtagctccatgccctgggtctttgtatgtgctat
ttctactgcttagagtgctattgcccctgaccaccac
gtggtcagcaacttctcttctgcgtctgtgtctatgg
tctatgattccagatgtcatcttcactaactaccctt
ctaatatgcccttccatcccacccgtcctcatcctta
ccccagccactctctatttggtggctctgttttattt
tcttcctagctcatcactctttgaaatgaacttattt
acttattcaattgcttctttcactagaatgaatgctc
catgagagcagggacctgctttatcttgctcgccact
gtattcacagtgcctagaactacgtctggcacatagt
aggtgctcaataaatatcgatcaaatgaaagaatgag
caaacgaacaaatgaacaacacgtgaggtaggcatca
tgattccatcaacagaggagaaaaccagacttaaagn
aatgaagtggnggagctgcatttgatcttgactgact
ccaacatccatgctcttgaccactgtgcatctccaga
gtgtaatgaacatactttacttttatattccaccaaa
ataacaaagccatgcccatgttagtagagagttaatc
gacagtgcccttaaaatatgcatgcacccagggtaca
actatgcatgctgccctgtgttttcagttggatccaa
atgaattgccgtaaacaaagaggggattcaatgtctt
tgactagtttgggatattttcctagtaaccaactttg
caaaataaagccactaatgacaaggagctttgttcta
cttctgcatcactcaactgtcaatttttatctcttgc
aagacttctaatctactagaacttttgtttttctgtg
atttctgaacagagaagactaatccaaaccctgtcat
tccagAGGAATGGAAAGCCCAATTCATTAAAACAGAA
AGGAAGAAACTCCTGAACTACAAGGCTCGGCTGGTGA
AGGACCTACAACCCAGAATTTACTGCCCCTTTCCTGG
GTATTTCGTGGAATCCCACCCAGCAGACAAgtatggc
tggatattttatataacgtgtttacgcataagttaat
atatgctgaatgagtgatttagctgtgaaacaacatg
aaatgagaaagaatgattagtaggggtctggagctta
ttttaacaagcagcctgaaaacagagagtatgaataa
aaaaaattaaatacaagagtgtgctattaccaattat
gtataatagtcttgtacatctaacttcaattccaatc
actatatgcttatactaaaaaacgaagtatagagtca
accttctttgactaacagctcttccctagtcagggac
attagctcaagtatagtctttatttttcctggggtaa
gaaaagaaggattgggaagtaggaatgcaaagaaata
aaaaataattctgtcattgttcaaataagaatgtcat
ctgaaaataaactgccttacatgggaatgctcttatt
tgtcagGTATATTAAGGAAACAAACATCAAAAATGAC
CCAAATGAACTCAACAATCTTATCAAGAAGAATTCTG
AGGTGGTAACCTGGACCCCAAGACCTGGAGCCACTCT
TGATCTGGGTAGGATGCTAAAGGACCCAACAGACAGg
tttgacttgaatatttacagggaacaaaaatgatttc
tgaattttttcatgtttatgagaaaataaagggcata
cctatggcctcttggcaggtccctgtttgtaggaata
ttaagtttttcttgactagcatcctgagcttgtcatg
cattaagatctacacaccaccctttaaagtgggagtc
ttactgtataaaataaactattaaataagtatctttc
aactctggggtggggggggagactgagttttttcaca
gtcctatataataattttcttatcctataaaatagga
gttcccgtagtggctcagcaatagcaaacccgactag
tatcgatgaggatgcgggttcgattcctggcccccct
cagtgggttaaggatctggcattgccgtgagctgtgg
tgtaggtggcagacacggctcagatcccacgttactg
tggctgtggcataggccagcagctccagctctgatta
gacccttagcctgggaacttccatatgctgtgggtgt
ggccttgaaaaaaaataaataaataagataattactc
aaatgttttccttgtctcagaaccttacttcaggata
aagagtgagaaagttttttttatgaagggccattatt
acagctcaaaaataagttgtcttcagcaagtagaaag
caataagcctgagagttagtgttcctatcagtgtaaa
tattacctcctcgccaatccccagacagtccatttga
acaattaacggtgccctgggagtacagttcagaaaca
ttaatgtggatgttccagacctgtatttttataagta
cttgtcttgagccggatggaaccatcattcctcacca
ttatttagaagtggactgtgactctgttggagatcag
ggcacacggttaccaaaagcacacccttctcctggcc
ttacctttgcaaagctggggtctgggacacagtcagc
tgattatacccttttactaacttcccacagctcaaat
ctggtcaattctccttcacaaatctcttaaaaatcca
tcactcacctccagcctcttctgctgtggccttgatt
cagcctctcacaatttttttttaaccagaattctggc
agtggcccctgacttgcctctgtgctcccagccccgc
tgtcctctgatccatcctccatgccagcctttttcaa
tctgctggtcacgattcattgatgggttaggaaatca
atggcatcacaactagcatttagaaaaaggaaatagg
cgttcccgccgtggcacagcagaaataaatccgacta
ggaaccataaggttgcgggttcaacccctggccttgt
tcagtgggttaaggatccggcattgccgtgggctgtt
ttgtaagtcacagacatggctctgatccggcattgct
gtggctctggcgtaggcctgcagcatcagctccaatt
agacccctatcctgggagcctccatatgctgcaagtg
cagccctaaaaaaaataaaaaaataaaaaaaaataaa
taaaagaagtagacaaattgtatagaacaaccctgag
tatgttgcctgagcacatataacaagggtaagtatta
tttcaggaaactctggtttcacagatactcttggcat
atggacccctagagtcctgatgtaaaatatattcttc
ctgggatcttaggcaagaagtttgaaagctccaactc
tgcactgctgccaaagaaatgatttttaagtgcaaaa
ctcttcccgttcccttccctgtataaaattccatagg
atctctccagtgcctctaggataaaggcagttttcat
tctctagttcaaggtgagagaagattttaattatttc
acgttttagtggggaattcaagagtctggcacctgac
atttgctgaactctctccattatccctctctagttcc
ccagacgcatcctatggtagaaattcgcaaactagag
tgagcgtcagagtaacccaaggaaactgggtaaatgc
agctccctgggctctaccccctgagattctgattcag
tagatctgaagcagagccctggaatatgcatatgcat
cattgtgtcacaccaagcattctgggtaatgagagtt
gatgttaggttctcagtagtaagacaagtatagagat
tccgggggactgagtgctcagctctgccttggggagg
agggagagggctaaagagaacaggagatggggacagg
gaatgctcaacctccaatcttaggcatttgagctatg
tcttaggggtcaggaggaggttaccaatatagtgatt
aagagattgaggttccagtcagagggatatgctggag
aaggggggtgaaaataatgtcataggtttggtgagtg
cagatactttgagttttttaatatttttattgaaata
tagttgatttacaatgctcttagtgagtacaattact
ttgaataagtgcatagatgtatgccattcttccagaa
atgatttattgagctcctttgggcatcatgctaagta
caggggaaacagctgtgaagaggtccttcccttatga
agtcattcatccccttcagtaaatgaaggtaaaggaa
aaggatgagacagggacgccgtgttggaccagggtca
gaaaggccttataagaccttgcctggagggcaaggaa
cttgcctgtgagtaaggagagcttgagaaagcgataa
agcaaagaaggaacattactgcattgtgttttagaaa
aaccatgtcctggggaagaactcctagagtcaggggg
gccagttgggagactgtgcttttttccaggaggagat
aagtgaggctgctggctgagatggagcaaggatttag
agaagcagatatgagattcatttagaagttagacatt
ttaggatctgacacataatttatcaccaaaaccagtg
catctctggctttgggccaccagttttggagaagtgg
aatgtagggacctaccattacctgccaatctttacta
cacagatgcctatttccctcctcatatttcctttctc
cagatcacgtcctattctattgccaggactcaagatt
ccaccttgcatgcagtgatccatcttcacactggatg
gacagctctagggatgtcagagcacactcttgtccat
actgctgactgggtctcctgtcagcccatctgtctat
cagctgtggtattattagtataataagagggctgtat
atgagagacacaaaattctaggtgtagctcaaagata
ggctagagttattcctatgtacaacaaatatttatgg
gaccccttctgtgtactgtcatggttgctgctttcat
catacttgtagtctaatggaggtgggggcagggcagg
aataagcggatgtccacaaaatcagtaagaccactta
tattcaacattttcataatttagttatttgagcccaa
agggtccacatccgtggtattccaacttttttttccc
cggacatggatctttatctttttttttttttcttttt
tgcggccagacctgcggcatatggaagttcccaggcc
aggggttgaatgggagttgcagctgcctggtctacac
cacagccacagcaaggtgggatctgagctgcatctgt
gacatacaccgcagctgaggtaacaccagattctgaa
cccactgaatgaggccagggatggaacccgtctcctt
atgaacactatgtcatgttcttcaccctctgagccac
aacgggaactccagacttcgtctttaaatgtattctg
acttggagagctatcacactaagcaattaacaggagc
tgacctggtttaggctggggtggggccctactcctca
atgttccctgaggcacatctgtgggacccctgggcat
catctatctgagcagccttagagctgctcatccagtt
gactgttgatgtagaagtgcaaacttctgccttcctt
atttgttgctttcttttttcattgttctctccccttt
gtgtctttaagCAAGGGCATCGTAGAGCCTCCAGAAG
GGACTAAGATTTACAAGGATTCCTGGGATTTTGGCCC
ATATTTGAATATCTTGAATGCTGCTATAGGAGATGAA
ATATTTCGTCACTCATCCTGGATAAAAGAATACTTCA
CTTGGGCTGGATTTAAGGATTATAACCTGGTGGTCAG
Ggtatgctatgaagttattatttgtttttgttttctt
gtattacagagctatatgaaaacctcttagtattcca
gttggtttctcaataagcattcattgagccttactga
ctgtcagacggagggcgtattggactatgtgctgaaa
caatcctttgttgaaaatgtagggaatgttgaaaatg
tagggaatgaaatgtagatccagctctgtttctcttt
tggaggattctttttcctccatcaccgtgtcttggtt
cttgtttgttttgggtttttgtgggtgttgtattgtg
ttgtgttggttatggcagtgacagctatttaaactgt
gaaacgggggagttcccgtcgtggcgcagtggttaac
gaatccgactgggaaccatgaggttgcgggttcggtc
cctgcccttgctcagtgggttaacgatccggcgttgc
cgtgagctgtggtgtaggttgcagacacggctcggat
cccgcgttgctgtggctctagcgtaggccagcggcta
cagctccgattggacccctagcctgggaacctccata
tgccgcaggagcggcccaaagaaatagcaaaaagaca
aaataaataaataaataaataagtaagtaaaataaac
tgtgaaacggggagttcccttcatggctcagcagtta
acaaacccagctaggatccatgaggatgtaggttcga
tccctggccttgctcagtgggttaagaatccagcgtt
gctgtgagctgtgatgtaggtcgcagatgcagcccag
atcctgcattgctgtggctgtggcgtaggctggcagc
tgaagctccgattcaacccctagcctgggaacatcca
tatgctgcaggtgtggccttaagaggcaaaaaaataa
aaaaataaaaaataaataaattgtgggacagacaggt
ggctccactgcagagctggtgtcctgtagcagcctgg
aagcaggtaaggtaaggactgcagctgggtaaggact
gaattgcaccaactgggaagtaagcctagatctagaa
cttaagttagccctgacatagacacacagagctcacc
agctaagtggttcagcttataagctggtcactgaaac
tgaggatgtccacaaaagcaaaataagtagcaacagg
cagcgggatgcaagagaaagaggaggcctaaaatggt
ctgggaatccctgccatacctatattttatcctactt
atatttagtgcctgaatgtgtgcctggagagcaaagt
ttagggaaagcatcgggaaatgcacagtattcatacc
cttaggaacaaagatcagttacctccagggtaaagac
tatttccaagtttaaatttcaacccctgaacattagt
actgggtaccaggcaacacttgccatcctcaaaatca
atgaatcctaaaattcaacctgggggtcagtgacagt
ctgtgacaaagtttttgctggtcagtaacgaaataag
tatgagcaccatctgagtatggtcaccaagatgtcaa
ctctctttcctttggacgaacattattccaagattag
gtcctttctatttttgaggtgtgaaaacatctttcct
ttcataaaataaaaggatagtaggtggaagaattttt
tttgttttttggtctttttgctatttctttgggccgc
ttctgcagcatatggaggttcccaggccaggggtcga
atcggagctttagccaccggcccacgccagagccaca
gcaacacgggatccaagccgcatctgcagcctacacc
acagctcacggcaatgccggatcgttaacccactgag
caagggcagggaccgaacccgcaacctcatggttcct
agtcggattcgttaaccactgcgccacgacgggaact
cctaatgatactcttttatatttagctactatgtgat
gatgagaaacagtccacattttattattttttagcca
atttgatatctcattactaagataatgataattttct
ctataaattttatttaagttagtgttatgaagtggtt
ttgctagtgtagaaggctaggatttgaattcagttca
agaaagaagagagggagggagggagagggatgggtag
agggatggggcagtgggagagagcaaagaggagagac
agtttttgtattaattctgcttcattgctatcattta
agggcacttgggtcttgcacattctagaattttctaa
ggaccttgaccgccagattgatatgcttcttcccttt
accatgttgtcatttgaacagATGATTGAGACAGATG
AGGACTTCAGCCCTTTGCCTGGAGGATATGACTATTT
GGTTGACTTTCTGGATTTATCCTTTCCAAAAGAAAGA
CCAAGCCGGGAACATCCATATGAGGAAgtaagcagga
ataccagtggaagtgcccctttcttccttccttccta
aataaacttttttattttggaacaactttagagttac
agaaaagttgcaaagatattatagacagtagtgttta
tatatatatataaatttttttttgctttttatgacca
cacctgtggcatatggaggttcccagtctaggggttg
aattggagctacagctgccagtctgtgccataaccac
agcaatgcaggatctgggccacgtctgtgacctacac
caaagctcacagctggattcttaacccactgagcaag
gccagggattgaacctgcatcctcgtggttcctagtt
ggattcgtttccgctttgccgcaatgggaactccaaa
ttattgttaatatcttactttactggggtacatttgt
tacaaccaatactctgatactgaaacattactgttaa
ctccgtacttgcttctttttgagtcatttgcaaagac
tggcttcttgacctgcttccttccaaacagctggcct
gcctatgctgttctcagacctgcaagcactgatctct
gccccccttgccttctctccagtggtgtctccttccc
caaacaaacccagtgtggctctggaaagggagttaag
tcaacataaaccaacacatattttgttgagctccaat
tttgagcaaatccctcaccacggcagacaggcatgat
gttaagaactagggctttggacacaaggtcaagacca
agaagggttcctcacccctactgattcagataaccaa
taatgaggctttgaatccctgtccaaaggttgttttt
tttcccttctattgagcttcttgccaccttatcagtt
ttttttatgacagtcaaatgacatgatatatgtgagc
atacatggtaatttttaattctatataaatgaatcac
taaataaattaggaggatatatagtccacctttaagc
gtattacacgtgtcacatgaatgtgtggcgacttaat
tgtagaggtttaaatgtagcttcctataatagatgtg
ttcctaaactacattttaatcattggacttgtatttt
tatgttagcacttgctgttgaagaaaagcctatgcca
aaagttcagtgaaaccaataatccactgccagctttc
tgagttaaaaaaaatccctgggttttcacacacagga
acaccctggaaacactcatttagagcaaaatgcatct
gataaggagttcctgttgtgcctcaactggttaagga
cctgacattctccatgagaatgtgagtttgatccccg
gccccactcgatgggttaaggatctggtgttgccaca
aactgcagctccgattcatctcctagcctagaaactt
ccacagcccagaatatgccacagaattcggctgttta
aaaaaaaaaagaaaaaaaaaagaatcataaatgtgtt
ggtttgttcaccaaatacatgataacttgctcttgcc
aagctcagcttcataaatattaagtcatttaatacag
cagccaccttatgaacagatattactatacttcccat
ttacagataaggaaaatgccatatttaaccaagagat
aataactttcccgaggtcttatagcaagtaaatcatg
gtgcaggggtttgaccacacgcagtctatctccagag
tctgtgtatttagccactgttttactttcaaatttaa
atttataaaacttctaaattatctgttaaccataatc
tttggaatttttaaaaccacgagttcctataaaatgt
ttcattgaaagtaagtcacttttccatagcttttgat
aatacatctgtaggataaagtaagccacagctctctt
gcagacttggtacaccctggggcaaagcatcatgcct
gtcacgtacatggtggtccttactttgactctcagtg
cttttattgcccaggaattttgtgagatttctagttg
ttgaggtttgtttaaagaggttatgccggtacttgga
agagctcttttcttgctacctggagccttctcatatt
tcctttttgaggagggacatgaattgcctttcaaact
cataaatatattttctagtacacaagtctccatcttc
cttagacgcatggctcctggagttctccatcctcctg
ctccactttgggtgggctcctctctgggtctgccacc
aatctgccacccagagacatccttgacccacttccag
accccaccatggcttcactttcttcgctttcctcctt
tgtggaaccttctgcttaagaatctgaggaagaaaat
ttgcacgtgagctaaactggaggtactttcctgcctg
gtcttgcacgatagcttggctgagcccatgatgctgg
gtggctgttactttccatggacacccgaaggcgttgc
tcctttggcttctagttgcatgcagtgttgcttatcc
caggctgatctttcttccactgtaggtgacttttaag
aattaagggattaatctatatctacaacaacaacaac
aaagaccttttcaagctgaggtagggctttctgtata
tgtttggagtggttatccagcagactttacttgaagg
caggggtcatatcctcaagtgctcataaacggaccac
agaaagatctcataattgggtggagctgggtggggac
cgtgtcatgtggccaggaaatgccagatgggaaggga
gtggcccttactgagctccagctgaactctgaatttt
ctagaaaactcagaaatctggatttttcatgtgtaat
acccagatttatagatgtggaaagctaattctttttt
tttttaagggactataggcaatgaactaagatctagg
ttgtatttggacaaggggtcatcagtttaagctgtgt
agttgagcgctcagctattgggctgagggacccctaa
atactgagacggggaggtccttgctctggggcatcac
aagtacactccctggtctcattcaaacacttttccta
caaaattgatcccatttcttcagtgcactgtctgaat
gcatttggcccagagccgtgctgaggcatagggaagg
ggtccacggtttcatggcatcgttttgtgctgtgtgt
ccctgctgtcgtccaggatacctacctctcctcctcc
tgcatctgaatgtccccccacagactctctgggattc
tacagcctctggcctgttcctcagacacctcttacct
gccagctttccagattcacattagttagccaaatcta
ctgccgtcagtgactcacttcatttcttcttctccga
ggcagttcagcccggtacagttgttttgtcaacactt
cagttgagtctggaagatgtgcatgggttatgcacga
gagcggtccatcattttgagctagaagtcctttctca
gcccagagacaagtcctcatctcctttacttcctgac
tcttcttcctctgcatccttccaagatatctctttct
ccagccaccacctaaatctcttcttttcccggggttc
cgtgctcaacccactcttcttcttaaatctgtggctg
ggtgaacgcatctgctggcaccacttctctgctaaag
actccaaaaatccataggtcctgcccggcctttgccc
acctctctccaacactgtccagctttagatgtagagc
taatccccccagagatatcattccctggatgtctaag
tcctttggtatctcactttcagcggttcaaaatcctc
ttacaactgttctttctccttttccatcttgattatt
ggcaacatgccagcctttcccctacccccagcagtga
gccaagctagaaacaagggcttaatcttcaatctttc
cttctccatccctaaacctaatgagtctccaagccct
tcccagtttacaccctaaatgttgctcaaaacatccc
ctagttcttccacgtgctctcctctatattgaaaggt
caagaaaggccatcttccctccactgtgaggaaatag
atcttgatactgcccctgagctgggcagtcctcgacc
tgacaaactgtgcagtgtttctaaatctctactggca
aaatgagagtgcctttgacctgtgttgcgatctcaga
tcacagtggatgtaattgttttataggaatggtgaac
gaaaaagaagtaaatccctaatgccaaactcctgatc
attctatgtcatttaatagcctgtcatttatgataaa
gtttcctctactggcattagcacaatacttctcagga
aaaaaaaatatgatgccagatactgaaaagctcctgg
gtaaacatgaacatgggtaccgataaaatggtgaagc
cagtccaatcttagagtgacttcccttcatgctactt
catgctcttttttttttttttttttaagaaaaacccc
ttttttttttctcacaccagtcacagaggagaccgag
gcttagcaaggttaaggtcacatgattagtaagtgct
gggctgaaactcaaaaccatctctgcttgtctcctaa
ccctgtgcacctctgactattcaacagATCCTGTGTC
AGGAGTTGGGATTCTTTGAAGgtaagggccttgacca
ccgaattaaggtaatcttgctctgtggcaggccttgt
tttcagtattttaagtacactggctcaggtaatcctc
acaacagccccaggaggaatgttctattacctccact
gtatagatgaggaacttgaggcacagaatggttgcca
aggtcacacagctatattgggggttcatacccagcca
tccaactctgtctgtactctctgccactctgcacccc
cagctcctgatccacttcctgtttccatccctcgatt
tctgctgcactcaggggcccctctccccctcggcctg
tgagatctgcttcagtaggcttttctccctgactcct
ccatccctgtccttacaggcagctgcttctctccggg
acacgaggggtccatacggacactctctactggctgg
gttgcgcctaactcgtgattcctcctctgtttcagAT
TCGGAGCCGGGTTGATGTCATCAGACACGTGGTAAAG
AATGGTCTGCTCTGGGATGACTTGTACATAGGATTCC
AAACCCGGCTTCAGCGGGATCCTGATATATACCATCA
TCTgtaagtccgaaaatgcctgtcgtgtgtgccttag
gctgctgcggaggaggccagggctatataagcagagt
cagtgactgactgtgccctgcagtgttgatggccatg
gagattccaccgttagagcttttttctttgttaacct
tgaaggcaaatctggttaggaagataactttcaaaga
gtcaccatctggacattcatgcccatgtgcttcaatc
ctgtatacaagcagtttagagtacagggaagggaagg
acattatgaaagggagagggtgtgtttggatccagca
gctccatcctcagaatttatctgaagacactgcaaaa
ttactaagaatcactatgacaagaatgaggatggggt
gatatggcaaagttgtgatcctggaagaccttcatct
cccatgttgcccaactctgaacatgaatttggtgaac
tagttggttaaggggatgatcctccaagtttctccct
ggttgagctccaaaaaccatgtaagtttctcatagca
aaaccgtataggtccttagggctttagttggaatatt
tgtgctgaaatgctggaaagccccatttgccattttt
gtatttgcaaaataatcatcaagaggggagaatgcat
tctttcatgaccactgaccctctgaaaaggtcaggaa
tttagtctgaagtaggcaagcctcctaccccgcttct
gccatgagcttgcacgcacaggcctgtcttgacattt
cttctttatagatttctttttgaatatcttgaaattg
ctttaaaaatatttaaagaatgtagaattatataaaa
taaaaaggaaataaccccacacctcccacaaaaccct
gtttcctgcctttctccacccactctccagggtaaca
cttggtaacagcatagttgtatcaccccaggcctatt
tttgagcatatcagcatttcaagaaatgtattttttc
tcaataaaacatcccttatagttgaggaggggaggtt
atcattcctgggttttgttttttttttttttttaatg
taatcctggtacatcggtaatttgcattttttattca
ttaatatctttggtatttctagtgttgggacacacag
gtcaacctcagtttttgggtttttttttttgtctttt
tgtctttctagggccacacctgcagcatatggacgtt
cccaagctaggagtctaatcagagctgtagccaccag
cctacgtcatagccatagcaacgtcagatccaagccg
tgtctgtgacctacaagcacagctcatggcaacaccg
gatccttaaccactgaacgaggccaggggatcgaaca
cacatcctcatggatcctagtcatgttcattaaccac
tgagtcatgatgggaactccaacttcaactatttaat
gtctgtaaaacattccatttggaaaccatttcatttg
taaagcaaaatgaaaacattttgttcattttcaacag
agttcgtagctgacttctgttctggaaaaaaggaaat
ggagcaaatttgagtgagaaagattcaaagataactt
ttcttttaaaaaaaattatatcttggaaacttctggg
ctattgattctgaagactatttttctatatactgttt
tgatagcaaagttcataaatgtgaaaggatcctgcga
tgaatcttgggaagcagtcatagcccaatatatcttt
gttgcttttaaaatgagatttagtttactaaatattt
ttctgatcataaaaataacacagatctaccgcagaaa
atttggaaaaaaaaaaacttttaaattcaaaaaacag
ttaaaccacaaatgatcccaccatccagagagcaatt
tgtactttggtgtctagttcatctttctttttctgtt
tacaagcacatataccacaagcattttttcaaaaaat
gaaaatgggataatactatacatacgtctgtacacct
gcatagttactgaacagtctttgatctaccctgtaag
tttctaacttttcattatttgaaatgatgttttggca
aagaaatatgtaggtgtgtctcgcacactttcataat
gatttcttaggataaatttcttaggataaattcataa
tgatttcttataataatccatactctgccaactgatc
ttcagggaagccaactcgccttctcagaaataacata
taacccatttacttgccctctcaccaatactaggtcc
taatgtttttgtgtacagattctatatttttacatac
aagaattccttaaagcaaggcatgtcacagaaaaata
gaaggaagacacaattgtcatgtttaaggactgcatt
ctgtaccaaaaatgctaagttaaatgaacatctgaaa
cagtacagaaacgctatctttcagggaaagctgagta
ccaggtactgaacagattttggcaaatacagcaggca
tggatgtttccaaaacatgtttttctactttatctct
tacagGTTTTGGAATCATTTTCAAATAAAACTCCCCC
TCACACCACCTGACTGGAAGTCCTTCCTGATGTGCTC
TGGGTAGAGAGGACCTGAGCTGTCCCAGgtaaagcat
cctgcaggtctgggagacactcttattctccagccca
tcacactgtgtttggcatcagaattaagcaggcacta
tgcctatcagaaaacctgacttttgggggaatgaaag
aagctaacattacaagaatgtctgtgtttaaaaataa
gtcaataagggagttcccatcgtggctcagtggtaac
gaaccctactagtatccattgaggacacaggttcaat
atctggcctcactcagtcggctaaggatccagtgatg
ccgtgagctgcagtgtaggccacagacgtggctcaga
tctggtgctgctgtggctatggtgtaggccggccccc
tgtaactccaattcgacccctaggctgggaacctaaa
aagaccccaaaaaagtcgcmatgaatagtgaatacat
ccagcccaaagtccacagactctttggtctggttgtg
gcaaacatacagccagttaacaaacaagacaaaaatt
atcctaggtggtcagtgggggttcagagctgaatcct
gaacactggaaggaaaacagcaaccaaatccaaatac
tgtatggttttgcttatatgtagaatctaaattcaaa
gcaaatgagcaaaccaattgaaacagttatggaagac
aagcaggtggttgtcaggggggagataaggggaggca
ggaaagacctgggcgagggagattaagaggtaccaac
tttcagttgcaaaacaaatgagtcaccagtatgaaat
gtgcaatgtgggaaatacaggccataactttataatc
tcttttttttttttgtcttttttgccttttctaaggc
tgctcccgtggcatatggaggttcccaggctaggagt
ccaaacagagctgtagctgccagcctacaccagagcc
acagcaacacgggaaccttaacccgctgagcaaggcc
agggatcgaacccgagtcctcacagatgccagtaggg
ttcattaaccactgagccacgacaggaattccagggt
ctgttgtgttcttaaaacacttccaggagagtgagtg
gtatgtcataagtaaacaataaatgttaaccacaaca
agcttatgaaataaacaggaaagccatatgacctaca
atcagtcattgggagaatccacaaaaggttgagcaga
ggatcaattccagctcacactccagttttagattctc
ccctgccttaaagcatcacagactacataatctgagc
tgaagaataaaaattaaaactcaccccagtgcaaaac
agaaatgaaaaagtattaaaacgaggttcatactgtt
gttcattagcaatatcttttattcacagGGGTGCCCA
ACAACATGAAAAAATCAAGAATTTATTGCTGCTACGT
CAAAGCTTATACCAGAGATTATGCCTTATAGACATTA
GCAATGGATAATTATATGTTGCACTITGTGAAATGTG
CACATATCCTGTTTATGAATCACCACATAGCCAGATT
ATCAATATTTTACTTATTTCGTAAAAAATCCACAATT
TTCCATAACAGAATCAACGTGTGCAATAGGAACAAGA
TTGCTATGGAAAACGAGGGTAACAGGAGGAGATATTA
ATCCAAGCATAGAAGAAATAGACAAATGAGGGGCCAT AAGGGGAATATAGGG
[0092]
14TABLE 13 SEQ ID NO. 49 TCTAATGCCTTGTGGAAGCAAATGAGCCACAGAAGCT SEQ
ID NO 49 GAAGGAAAAACCACCATTCTTTCTTAATACCTGGAGA
GAGGCAACGACAGACTATGAGCAG gcaagtgagagggggctttagctgtcagggaaggcgg
agataaacccttgatgggtaggatggccattgaaagg aggggagaaatttgccccagcaggtagc-
caccaagct tggggacttggagggagggctttcaaacgtattttca
taaaaaagacctgtggagctgtcaatgctcagggatt ctctcttaaaatctaacagtattaatct-
gctaaaaca tttgccttttcatag CATCGAACAAACGACGGAGATCCTG- TTGTGCCTCTCA
CCTGCCGAAGCTGCCAATCTCAAGGAAGGAATCAATT
TTGTTCGAAATAAGAGCACTGGCAAGGATTACATCTT ATTTAAGAATAAGAGCCGCCTGAAGGCA-
TGTAAGAAC ATGTGCAAGCACCAAGGAGGCCTCTTCATTAAAGACA TTGAGGATCTAAATGGAAG
gtactgagaatcctttgctttctccctggcgatcctt
tctcccaattaggtttggcaggaaatgtgctcattga
gaaattttaaatgatccaatcaacatgctatttcccc cagcacatgcctaactttttcttaagct-
cctttacgg cagctctctgattttgatttatgaccttgacttaatt
tcccatcctctctgaagaactattgtttaaaatgtat tcctagttgataaacagtgaaacttcta-
aggcacatg tgtgtgtgtgtgtgtgtgtgtgtgtgtttaccagctt
ttatattcaaagactcaagcctcttttggatttcctt tcctgctctctcagaagtgtgtgtgtga-
ggtgagtgc ttgtccaaacactgccctagaacagagagactttccc
tgatgaaaacccgaaaaatggcagagctctagctgca cctggcctcaacagcggctcttctgatc-
atttcttgg aagaacgagtgctggtaccccttttccccagcccctt
gattaaacctgcatatcgcttgcctccccatctcagg agcaattctaggagggagggtgggcttt-
cttttcagg attgacaaagctacccagcttgcaaaccagggggatc
tggggggggggtttgcacctgatgctcccccactgat aatgaatgagggattgaccccatctttt-
caagctttg cttcagcctaacttgactctcgtagtgtttcagccgt
ttccatattaggcttgtcttccaccgtgtcgtgtcgt caatcttatttctcaggtcatctgtggg-
cagtttagt gcgaatggactcagaggtaactggtagctgtccaaga
gctccctgctctaactgtatagaagatcaccacccaa gtctggaatcttcttacactggcccaca-
gacttgcat cactgcatacttagcttcagggcccagctcccaggtt
aagtgctgtcatacctgtagcttgcttggctctgcag atagggttgctagattaggcaaatagag-
ggtgcccag tcaaatttgcatttcagataaacaacgaatatatttt
tagttagatatgtttcaggcactgcatgggacatact tttggtaggcagcctactctggaagaac-
ctcttggtt gtttgctgacagactgcttttgagtcccttgcatctt
ctgggtggtttcaagttagggagacctcagccatagg ttgttctgtcaccaagaagcttctgcaa-
gcacgtgca ggccttgaggtcttccgacttgtggcccggggactct
gctttttctctgtccttttttctccttagtgggccat gtcctgtggtgttgtcttagccagttgt-
ttaagggag tgttgcagctttatgattaagagcatggtctttcctt
gcaaactgcttggtttagaagcctggctccaccactt agcggctctgtgacctcggacacatttc-
ttagccttt ctgggcctcgctcttcttcctcataaagtgaaaatga
aagtagacaaagccttctctgtctggctactgagagg atggagtgatttcatacacataaagcac-
ttaaaataa tgtctggcatatgatacatgctcaataaatgtcactt
acatttgctattattattactctgccatgatcttgtg tagcttaagaacagaggtctttacagga-
attcaggct gttcttgaatctggcttgctcagcttaatatggtaat
tgctttgccacagactggtcttcctctccttcaccca aagccttagggggtgaacgatcccagtt-
tcaacctat tctgttggcaggctaacatggagatggcaccatctta
gctctgctgcaggtggggagccagattcacccagctt tgctcccagatacagctccccaagcatt-
tatatgctg aaactccatcccaagagcagtctacatggtacactcc
cccatccatctctccaaatttggctgcttctacttag gctctctgtgcagcaattcacctgaaat-
atctcttcc acgatacagtcaagggcagtgacctacctgttccacc
ttcccttcctcagccatttttcttctttgtacataat caagatcaggaactctcataagctgtgg-
tcctcattt tgtcaatctaatttcacagcctcttggcacatgaagc
tgtcctctctctcctttctgcctactgcccatgagca gttgtgacactgccacatttctccttta-
acgacccag cctgctgaatagctgcatttggaatgttttcaatttt
tgttaatttatttatttcatctttttttttttttttt tttttttttttttttagggccgcaccca-
tgggatatg gaggttcccaggctagggatccaatgggagctgtagc
tgctggcctacaccacagccacagcaatgcacaattc gagccaatctttgacctacaccagagct-
cacggcaac actggattcttaacccactgattgaggccagggatca
aactctcgtcctcatagatacgagtcagattcgttaa cctctgagccatgatagttgttagttac-
tcattgatg agaaaggaagtgtcacaaaatatcctccataagtcga
agtttgaatatgttttctgccttgttactagaaaaga gcattaaaaattcttgattggaatgaag-
cttggaaaa aatcagcatagtttactgatatataagtgaaaataga
ccttgttagtttaaaccatctgatatttctggtggaa gacatatttgtctgtaaaaaaaaaaaat-
cttgaacct gtttaaaaaaaaaacttgactggaaacactaccaaaa
tatgggagttcctactgggacacagcagaaatgaatc taactagtatccatgaggacacaggttt-
gatgcctgg cctcgctaagtgggttaaggatatggtgttgctgcag
ctccaattcaacccctatcctgggaacccccatatgc caccctaaaaagcaaaaagaaaggtgct-
gccctaaaa agcaaaaagaaagaaagaaagacagccagacagacta
ccaaatatggagaggaaatggaacttttaggccctat ctccaactatcacatccctatcaccgtc-
tggtaagaa atggaaaaaatattactaagcctcctttgttgctaca
attaatctgattctcattctgaagcagtgttgccaga gttaacaaataaaaatgcaaagctgggt-
agttaaatt tgaattacagataaacaaattttcagtatatgttcaa
tatcgtgtaagacgttttaaaataattttttatttat ctgaaatttatatttttcctgtatttta-
tctggcaac catgatcagaaatctttaaacaatcaggaagtctttt
ttcttagacaaatgaaaatttgagttgatcttaggtt tagtacactatactaggggccaagggtt-
atagtgtga ctattaaatcacagataatctttattactacattatt
tccttatactggccccacttggatcttacccagctta gcttttgtatgagagtcatccttaaaga-
tgactttat tctttaaaaaaaaaaacaaattttaagggctgcaccc
atagcatatagaagttcctaggctagcggtcaaatta gagctgcagctgccagcctatgccacag-
ccacagcaa tgccagatctgagctgcatctgtgacctacactgcag
cttgcagcaatgctggatccttaacccattgaacaat gccagggattgaacacacatcctcatgg-
atactgctc aggttcctaacctgctgagccacagttggaactccaa
agcagactttattctgatggctctgctgatctctaac acgttattttgtgccatggtgtttatct-
tcactttac tcaagtcagggaaacacgaagagtctcatacaggata
aacccaaggagaaatgtgcaaagtcacatacaaatca aactgacaaaaatcaaatacaaggaaaa-
aatatcttc actttcaaaatcacctactgatgatgagtttatattt
ccttggatatttgaatattagctatttttttcctttc atgagttttgtgttcaaccaactacagt-
cgtttactt tgatcacagaataatgcatttaagccttaaatagatt
aatatttattttcaccatttcataaacctaagtacaa tttccatccag
GTCTGTTAAATGCACAAAACACAACTGGAAGTTAGAT GTAAGCAGCATGAAGTATATCAATCCTC-
CTGGAAGCT TCTGTCAAGACGAACTGG gtaaataccatcaatactgatcaatgttttctgctgt
tactgtcattggggtccctcttgtcaac- ttgtttcca
atctcattagaagccttggatgcattctgattttaaa
ctgaggtattttaaaagtaaccatcactgaaaattct aggcaagttttctctaaaaaatcccttc-
attcattca tttgttcagtaagtatttgatgagaccttaccatgtg
taaacattgcactaggtattaagaaatacaaagatgg ataagatagagtcggcgtaaatgagatg-
atataatga gacgttataatgaaactcacaattccagttgggaaat
aaagtccttcaaattccatgactctttctggcacacg ttagaggctacagcttctgtgtgattct-
catgctggc tccacttccactttttccttcttcctactcaagaaag
cctatagaaatatgagtaagaagggcttaatcatagg aataaatttgtctctgttctaagtgatt-
aaaaatgtc tttatcagtataaaaagttacttgggaagattcttaa
aactgcttttacacactgttctagaatgactgttata taaataaaaaagtagatttgatctaaca-
caattaaat gacctttggaaatattgactaattctcaccttgcccc
tcaaagggatgcctgaaccatttccttcttttgccag aaagcccccaccctttgtctgttgacct-
agcctagga aatcttcagatcacgttgttagcacgaactggttaca
tgtgctgtacaaatactatttaattcatctgattaaa aaaaaagagataagaagcaaaagtttga-
ctatcttaa actgtttgcgtaggtgagaggacaattgaccatctac
tttatgagtatgtaacccagaaacttaaagctcctta agggagctaagtcttttggataagacct-
atagtgaga ccttttagcaaaatggttaagactgaatggagctcac
tagcgtgggttcatatcctgatgctcaaacacgcaat taaatgactttaggtgggttagtctctg-
ttccttagt ttcctcaatgggagataatattggtagtagcgatttt
actgggttgttgaaagaacatctgttaaatgttcaga acgtgttacgacagagtacagagtaatg-
atttgcttg tatatgtatgactcaaatagtctgccatatgccttgt
gactgggtcctgtggagcaggaaggagggatttccca cccagcagaaagttgggtaaactggaaa-
atagactga ggccaggaaatgatgcaaagcgttgatgttcactgcc
acggcaggtgaagggcagggccagagttgtcagtagg gtcaggggaggactggaaataaccaaga-
cccactgca cttttcagcctttgctccagtaaggtaatgttgtgag
agtagaaaattttgttaacagaacccacttttcagta cagtgctaccaaactgtagtgatttcat-
accacatcc caagaaagaaaaagatggctcaatcccatgtgagctg
agattatttggttttattgttaaataaatagcattgt gtggtcatcattaaaaaaggtagatgtt-
aggaaagta gaaggaagaagactctcacctacattttcatcactgt
tttggtatctgccagttgtcaccttggtccccttccc cgcctctcccctgcctcctcttcctcct-
tctcctttt tttggaatacaattcaggtaccataaaatttaccctt
ttagagtgtttgactcaatggtttttagtattttcac atgttgtgctattactatcactatataa-
ttccaggtc attcacatcaccccccaaagaaaccttctaactatta
gcagtccattcccttcttccctcagcccctggcaacc actaatctacttactgtctccatggatg-
ttcctatat tgaatcaagctagcataaaccccacttgctcatggtc
ataattcttttttatagtgctaaattacatttgctaa tattcaattaaggatttctatgtccata-
ttcataagg aatattggtgtgtagttttctctttgtgtgatatctt
tgtctggttgggggatcagagtaataattactgctct catagaatgaattgagaagtgttccctc-
cttttctat ttattggaagagtttgtgaagtatattggtattgatt
cttctttaaacatttggtcagattcaccagtgaagcc atctgggccatggctaatctttgtgaaa-
agttttttg attactaattaaatctctttaatttgttatgggtctg
ctcctcagacgttctagttcttcttgagtcagttttg ttcatttgtttcttcctaggactttctc-
cctttcatt tggattatttagattgatagtaatatcccccttttaa
ttcctggctgtagtaatttgggtcttttctctttttt cttggtcagtttagctaaaggtttgtaa-
ttgtattaa tcttttcaaataactaacttttttgttttgtttgttt
tttgttttttgttttttgttttttgtttttttttgct ttttaaggctgcacctgaggcatatgga-
agttctcag gctagaggtctaatcggagctacagctgctggcctat
accacaaccatagcaatgccagattcaagctgcatct gcgacctacaccacaactcggccaggga-
tcacacccg caacctcatggttcctagtcggatttgttaaccactg
tgccacgacgggaactcccgcccattttttttaacac ctcatactttaacataaagatgggcttc-
acatggact gatagctcaaatgaggaaggtaagactatgaaagtaa
tggaagaaatgtagactatttttgtgacctagagatt actgatacttcttgacttttcaaacaat-
acttcaaaa gtacagcccaaagggaaaaaagaaagaaaaaagaaac
acacatatacacaaacctagtgaataagatatcatcg atacactacagatttctatgaactggaa-
gaccccatg gacaaagttaaagaacatatgatagtttgagtgatta
ttttgcaatatttacaaccaatgagggaatattatcc agcttataggaggaagtaatgcaaatcg-
acaagaaaa agataggaaacccaatataaaaattaagaaaatacaa
aaattaagaaaggatatgaactagcattttacaaaag aaaaatctccaaaagtcaatcagcacat-
gaaaatatg ctcaaacctattaattattagaaaactacagactgaa
gcaatgaggtgctttactttacatctttttgactgat aaaaagttagaaacaaaggtgatatcaa-
atgtcaggg ataaaaggatatagaaatcgtcatgcctgtggtggga
gtatggccggtgcagtcatgtgggaaggtaatctgac agtggttaggcagagcaggtttatgaat-
acactgtgg cccatcaatcccacgcctgtttatgtaccaaagaaat
cctgttgtggcagaatctatgggtccacccctgggag catgaattaataaaatgtggcaccaggg-
tgtgtgaaa ctccagctagagatgagatgtccacatggcaacatga
atgcatcttagaaacatagatttgagtgaaaaagagt aagaaacagccgggaaacccaataccat-
ttataaaaa ttaaagatgcacacatacaatgtagtaaatattttgc
atgaactttcaaatggttgcctacagggggggagagt aaagaagagtagaaaacaaagataaagg-
gagtaagta agtagctctgcctggactgaatataatgtgtcatgaa
ctgagaaatatggttaacataatcctctaacttgagg tcctaaatgaatgaatgagtccactatt-
catttaccc attctttaatgtgtattgcattataatccattttttt
agaaccaacgaattttgttcccataactactaatcag cctgccttttctccctcattcccttatc-
agctcaggg gcattcctagtttttcaaacgttcctcatttgaacca
aaaatagcatcattgtttaaattatacttgttttcaa atacgatgcttatatattccaagtgtgt-
ttgcccatt ttcttaggtggtagaaatttttcattctacttttcta
tctactcagattttcccgttggaattatttccattgc tattaaacttagaagtcccccctgtgat-
atgccattt ttttcatactttttaagcacttggttgcttttctttg
tgtctttaagcacctagaatacttataaccattgcac agcactgtgtatcaggcagcccttcctc-
ttccactaa tttatggtccttctcttagactatattaaactgttat
ttaattaggatcctctcttcgtccttatgatttaatt attatagttttctaatatgtttttatta-
taattcctc ttcattattcctccctattaaaaattttaatgaattc
catttgtttgttcttctagttaaatattaagtcataa tccaaataacttagatgtcattagttta-
tgtggtcaa agtaaggataccacatctttatagatgcaggcagttg
gcagatgtcatgattttcttcagtgcataaatgcaat ttatctttgagcaaggggcataaaaact-
tttatggta ttggctttgaaataatagttaagaactgcagactcag
tttttcctgcttttcttgaaaaagaacacttctaaag aaggaaaatccttaagcatggatatcga-
tgtaatttt ctgaaagtctcctgtaattccttgggatttttgttgt
tgtttgttggtcggtttttttgggtttttgtttgttt gttttgttttgttttgttttgcttttag-
ggctgcacc tgtggcatatggaagttcccaggctaggggtccaact
ggagctacagctgccagcctactccacagccacagca acatgggatcctagctgcatctgtgacc-
taaccacag ctcttggtaatgccagattgttaacccactgagcaat
gccagagatcgaatctgcctcctcatggacactagtc agattagtttctgctgagccacaatggg-
aattcccaa ttccttgtatttttgaactggttatgtgctagcatat
aattttgtttcttgaatctttgtgggttttttttttt ttttttttttgtctcttgtctttttaag-
gctgcaccc acagcatatggaggttcccaggctagaggtcaaattg
gagctacagctgccagcctacacaacaactgcagcaa agtggggcccaacttatatgacagttcg-
tggcaatgc cggattcctaacccactgagcagggccagggatcgaa
cctgagtttccagtcagtttcgttaaccactgagcca tgatagtaactcctgtttgttcagtctt-
gaacctcct ttttaattctttattccttgagggtgaaataattgcc
ataataatactatcatttattacatgccttctctgtg ctaggcatagtgacactttaggatttat-
tatatcact taatccctacaacaactctgcaaagtatgtatcataa
tcctatttgacagatcaggaaattgcagcccaggatg cagataatatgcatccatcacaagtgac-
tagatatag tccctctgctattcagcagggtctcattgcctttcca
ttccaaatgcaatagtttgcatctattgtatatgtgt tttggggtttttttgtcttttttttttt-
ttttgtctt ttctggggcctcacccttggcataggtaggttcccag
gctaggggtcaaattgaagctgcagctgccagcctac accacagccacagcaactcgggatctga-
gcctcatct gcaacctacaccaaagctcacggcaacaccggatcct
taacccactgagtgaggccagagatcaaaccggcaac ctcatggttcctagtcggattcattaac-
cactgagcc acgatgggaactccctaaatgcaatagtttgctctat
taaccccaaactcccagtccatcccactccctcctcc tccctcttggcaaccacaagtctgttct-
ccatgtcca tgattttcttttctggggaaagtttcatttgtgccat
ttttcattttacgggtaatttttacttcagtttcttc cactagcagttgtcttaaagtgagtata-
attaatatt catttggaaaatgtaagcaaaacattttttaaagggc
catgcccacagcatatgaaagtttctgggccaggggt tgaatccaggctccaagttgcagctgtg-
ccctacact gcagctgggcaatgctggatcctttaacccactgtgc
ccggctagggatcaaacctgcatttccacagctaccc gagccattgcagttggattcttaaccca-
ctgcactac agtgggaactcccacaaaacattttttaatgtccttt
gaataaagtaggaaagtgctcgtctttgagggcaggg cggcaatgccatttccacaaggtttgct-
ttggcttgg gacctcatctgctgtcatttagtaatgaataaaattg
ctgacagtaataggattaactgtgtgtggagatagcc agggttagagataaaaacactggagaag-
tcaaataag ttgctcgaggtcctctagctaataagctattaagtgg
gagagtgagggctagaaacaggccatctgtctcccaa gcacatgtccattagtggtttgctgata-
gccttccag aacaacagagaggactctcaaacatggtcttgcctcc
ctccaattgatcccctccatgtgcctcacagcgggtc tttctaaaattaagttctgattttaatt-
ctcccttgc tatagcacttaggtatggctttcagccgtgcaataaa
aagcaggcaagagtggctcaatcatataggaggttgt ttttcttagatcccaagcaggtaatcct-
gggcattat ggttgttctgcgtttatcaaggagccaaattctctat
cacctcctgttctatcctcctcagtatctggctctat tcttcagcatctcaagatggcttgtgct-
cctccaagc atggcagtcaaattccacacaagagggggaaatatga
agggcagacagtgctggtctcctgagctgtccctctt tgtcggggaaaatgtattccttcaagtc-
ccgtgagac ttctgaagtagacgtctgcttacgtctcacccaccag
aactatgtaaactgcacatagtgctaggtctacatag ccactcataactgccagggggtgggaaa-
tctttaaat aggtgtaccaccacacaattaggatgctaatagtaag
ggagaaggagagaataggttttgcgcaagccaccagc atgcctgccacaattgcttaaaattctt-
cattgaccc ctcattgccacaggatgaaatccaaacgccttcttag
ttgggaatctgacctacctgtctctcccacctggttc agacaccattctccttggtcataaaatt-
ccagtcatt tgtgaacatccagctcccccatgcctccatgcctttg
cacatgctgttcttttatcttttatgttgtcctttta tcttttatccaaaagagatatcccatca-
tcacatctc ttttgtcagcccccaaatactttgtctttcaagttca
gctggaggattacctcctatttgaaatcagctttgtc tcttacaaccaaacaaggttttccttcc-
gagacactc ccacagcaccttgaactcatctctatcaatcattcat
ttgattgtaatgaagttgttggtggtatgcctgtgtc tctgacacatctgcgatctcatgagttc-
cttaagtgg aatgtgaatagcgggatgaacagtattggtcttcagc
cctcatctctgcagatgttgcttgacccaaatgagcg ttgccttttattttgattttgctttgat-
ttgtctact ccatgtacttgagccatgcatttctgtcttagcgatg
ctttttaaaagtcattttttggttgattatccagatt tgtccacctttgcttctag
TTGTAGAAAAGGATGAAGAAAATGGAGTTTTGCTTCT
AGAACTAAATCCTCCTAACCCGTGGGATTCAGAACCC AGATCTCCTGAAGATTTGGCATTTGGGG-
AAGTGCAG gtaaggaaatgttaaattgcaatattcttaaaaacac
aaataaagctaacatatcaatttatatatatatatat atatatatttttttttttttttacatct-
tatattacc ttgagtattcttggaagtggctagttaggacatataa
taaagttattctgaagtctttttttttctttttccat ggtgagcagtggcttgatgtggatctca-
gctcccaga cgaggcactgaacctgagccgcagtggtgaaagcacc
aagttctagccactagaccaccagggaactccctatt ctaaattcttgagcacattatttaggaa-
cctcaggaa cttggcaggattacaggaaatatatctagatttaaaa
aaaaatcttttaacagaggtcccaaaggagagtcatg cacagctatgggaggaagttcagaaact-
gcccttgct accagatcactgtcagataaaatggccagctacatgt
ttctgcacattgccctaagatctttacaaacttttct gtgcatttttccacttttaaaagaaaat-
ttcggggtt cctgttgttgctcagtggttaacgaacccaactagta
tccatggggacaggggttcgagccctggcctcactca
gtgggttaagaatctggcattgctgtgg-
ctgtggcgt aggctggcggctacagctcagattggacccctagcct
gagaacctccatatgccgcaggtatggccctaaaaaa aaaaaaaaagagagagagagaatttcct-
ccagaaaaa acactttggtagtttgggagaagtaaacaaccaaaaa
ttaatttttctggagtattcgggaagcttgtaaaaat gggctcttacttttttgaggagacaaat-
gggaaccta cccagaagaggcacaatcacctgcatttgatttcttg
acctctccctaccttctttgctggctttccacatttg gatttctgtgaccttatctctgctcctt-
ggtgttttc atttttcctgtggacgtgccagactatgggaagggag
taaggcgttgatttagaatcctgtagtctctgcctgt ctctagtcattgttttcacccttctcaa-
aggaccttg acatcctgagtgagtccgcaagtaatttaggggagaa
gccttagaagccagtgcagccaggctacatgactgtg tccacccactggaaccagtcatttttat-
acctattca cagcccccctaccatttaaatccccagaggtctgcca
taacatctgtaactccctttcctggtaaattgtgttc taaaagactggtaacaaaagatattctg-
tggtacaga gcataattaaatacctgggagctgatttgagtggggt
aaatcaactggtttgacccctaaaacccaccatgagc atttctgttctaataaagtaatgcccgt-
gctgggaat tgtgttctacggaaatgctcctgctgtgtctttcttg
agtcctgtgtcattgaacatgcttaggagcaaaggtc ccccatgtggcttgtctgctaaccagcc-
cagttcctt gttctggctggtaatgatccgatcatctgaatctcac tgtcttccaacag
ATCACGTACCTTACTCACGCCTGCATGGACCTCAAGC
TGGGGGACAAGAGAATGGTGTTCGACCCTTGGTTAAT CGGTCCTGCTTTTGCGCGAGGATGGTG-
GTTACTACAC GAGCCTCCATCTGATTGGCTGGAGAGGCTGAGCCGCG
CAGACTTAATTTACATCAGTCACATGCACTCAGACCA CCTGAG
[0093] SEQ ID NO. 49 represents contiguous genomic sequences
containing Intronic sequence 5' to Exon 4, Exon 4, Intron 4, Exon
5, Intron 5, Exon 6, Intron 6, Exon 7, Intron 7 and Exon 8 (Table
13). Further, nucleotide sequences that contain at least 1750,
1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800,
2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900,
4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000,
5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, 11000,
12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000, or 20000
contiguous nucleotides of SEQ ID NO. 49 are provided, as well as
nucleotide sequences at least 80, 85, 90, 95, 98, or 99% homologous
to SEQ ID NO. 49.
15TABLE 14 SEQ ID NO. 50 AGGAATGGAAAGCCCAATTCATTAAAACAGAAAGGAA SEQ
ID NO. 50 GAAACTCCTGAACTACAAGGCTCGGCTGGTGAAGGAC
CTACAACCCAGAATTTACTGCCCCTTTC- CTGGGTATT TCGTGGAATCCCACCCAGCAGACAA
GTATGGCTGGATATTTTATATAAC- GTGTTTACGCATA
AGTTAATATATGCTGAATGAGTGATTTAGCTGTGAAA
CAACATGAAATGAGAAAGAATGATTAGTAGGGGTCTG GAGCTTATTTTAACAAGCAGCCTGAAAA-
CAGAGAGTA TGAATAAAAAAAATTAAATAC
gtatggctggatattttatataacgtgtttacgcata agttaatatatgctgaatgagtgattta-
gctgtgaaa caacatgaaatgagaaagaatgattagtaggggtctg
gagcttattttaacaagcagcctgaaaacagagagta tgaataaaaaaaattaaatacaagagtg-
tgctattac caattatgtataatagtcttgtacatctaacttcaat
tccaatcactatatgcttatactaaaaaacgaagtat agagtcaaccttctttgactaacagctc-
ttccctagt cagggacattagctcaagtatagtctttatttttcct
ggggtaagaaaagaaggattgggaagtaggaatgcaa agaaataaaaaataattctgtcattgtt-
caaataaga atgtcatctgaaaataaactgccttacatgggaatgc tcttatttgtcag
GTATATTAAGGAAACAAACATCAAAAATGACCCAAA
TGAACTCAACAATCTTATCAAGAAGAATTCTGAGGT GGTAACCTGGACCCCAAGACCTGGAGCCA-
CTCTTGA TCTGGGTAGGATGCTAAAGGACCCAACAGACAG
gtttgacttgaatatttacagggaacaaaaatgattt ctgaattttttcatgtttatgagaaaat-
aaagggcat acctatggcctcttggcaggtccctgtttgtaggaat
attaagtttttcttgactagcatcctgagcttgtcat gcattaagatctacacaccaccctttaa-
agtgggagt cttactgtataaaataaactattaaataagtatcttt
caactctggggtggggggggagactgagttttttcac agtcctatataataattttcttatccta-
taaaataat taggagttcccgtagtggctcagcaatagcaaacccg
actagtatcgatgaggatgcgggttcgattcctggcc cccctcagtgggttaaggatctggcatt-
gccgtgagc tgtggtgtaggtggcagacacggctcagatcccacgt
tactgtggctgtggcataggccagcagctccagctct gattagacccttagcctgggaacttcca-
tatgctgtg ggtgtggccttgaaaaaaaataaataaataagataat
tactcaaatgttttccttgtctcagaaccttacttca ggataaagagtgagaaagttttttttat-
gaagggcca ttattacagctcaaaaataagttgtcttcagcaagta
gaaagcaataagcctgagagttagtgttcctatcagt gtaaatattacctcctcgccaatcccca-
gacagtcca tttgaacaattaacggtgccctgggagtacagttcag
aaacattaatgtggatgttccagacctgtatttttat aagtacttgtcttgagccggatggaacc-
atcattcct caccattatttagaagtggactgtgactctgttggag
atcagggcacacggttaccaaaagcacacccttctcc tggccttacctttgcaaagctggggtct-
gggacacag tcagctgattatacccttttactaacttcccacagct
caaatctggtcaattctccttcacaaatctcttaaaa atccatcactcacctccagcctcttctg-
ctgtggcct tgattcagcctctcacaatttttttttaaccagaatt
ctggcagtggcccctgacttgcctctgtgctcccagc cccgctgtcctctgatccatcctccatg-
ccagccttt ttcaatctgctggtcacgattcattgatgggttagga
aatcaatggcatcacaactagcatttagaaaaaggaa ataggcgttcccgccgtggcacagcaga-
aataaatcc gactaggaaccataaggttgcgggttcaacccctggc
cttgttcagtgggttaaggatccggcattgccgtggg ctgttttgtaagtcacagacatggctct-
gatccggca ttgctgtggctctggcgtaggcctgcagcatcagctc
caattagacccctatcctgggagcctccatatgctgc aagtgcagccctaaaaaaaataaaaaaa-
taaaaaaaa ataaataaaagaagtagacaaattgtatagaacaacc
ctgagtatgttgcctgagcacatataacaagggtaag tattatttcaggaaactctggtttcaca-
gatactctt ggcatatggacccctagagtcctgatgtaaaatatat
tcttcctgggatcttaggcaagaagtttgaaagctcc aactctgcactgctgccaaagaaatgat-
ttttaagtg caaaactcttcccgttcccttccctgtataaaattcc
ataggatctctccagtgcctctaggataaaggcagtt ttcattctctagttcaaggtgagagaag-
attttaatt atttcacgttttagtggggaattcaagagtctggcac
ctgacatttgctgaactctctccattatccctctcta gttccccagacgcatcctatggtagaaa-
ttcgcaaac tagagtgagcgtcagagtaacccaaggaaactgggta
aatgcagctccctgggctctaccccctgagattctga ttcagtagatctgaagcagagccctgga-
atatgcata tgcatcattgtgtcacaccaagcattctgggtaatga
gagttgatgttaggttctcagtagtaagacaagtata gagattccgggggactgagtgctcagct-
ctgccttgg ggaggagggagagggctaaagagaacaggagatgggg
acagggaatgctcaacctccaatcttaggcatttgag ctatgtcttaggggtcaggaggaggtta-
ccaatatag tgattaagagattgaggttccagtcagagggatatgc
tggagaaggggggtgaaaataatgtcataggtttggt gagtgcagatactttgagttttttaata-
tttttattg aaatatagttgatttacaatgctcttagtgagtacaa
ttactttgaataagtgcatagatgtatgccattcttc cagaaatgatttattgagctcctttggg-
catcatgct aagtacaggggaaacagctgtgaagaggtccttccct
tatgaagtcattcatccccttcagtaaatgaaggtaa aggaaaaggatgagacagggacgccgtg-
ttggaccag ggtcagaaaggccttataagaccttgcctggagggca
aggaacttgcctgtgagtaaggagagcttgagaaagc gataaagcaaagaaggaacattactgca-
ttgtgtttt agaaaaaccatgtcctggggaagaactcctagagtca
ggggggccagttgggagactgtgcttttttccaggag gagataagtgaggctgctggctgagatg-
gagcaagga tttagagaagcagatatgagattcatttagaagttag
acattttaggatctgacacataatttatcaccaaaac cagtgcatctctggctttgggccaccag-
ttttggaga agtggaatgtagggacctaccattacctgccaatctt
tactacacagatgcctatttccctcctcatatttcct ttctccagatcacgtcctattctattgc-
caggactca agattccaccttgcatgcagtgatccatcttcacact
ggatggacagctctagggatgtcagagcacactcttg tccatactgctgactgggtctcctgtca-
gcccatctg tctatcagctgtggtattattagtataataagagggc
tgtatatgagagacacaaaattctaggtgtagctcaa agataggctagagttattcctatgtaca-
acaaatatt tatgggaccccttctgtgtactgtcatggttgctgct
ttcatcatacttgtagtctaatggaggtgggggcagg gcaggaataagcggatgtccacaaaatc-
agtaagacc acttatattcaacattttcataatttagttatttgag
cccaaagggtccacatccgtggtattccaactttttt ttccccggacatggatctttatcttttt-
ttttttttc ttttttgcggccagacctgcggcatatggaagttccc
aggccaggggttgaatgggagttgcagctgcctggtc tacaccacagccacagcaaggtgggatc-
tgagctgca tctgtgacatacaccgcagctgaggtaacaccagatt
ctgaacccactgaatgaggccagggatggaacccgtc tccttatgaacactatgtcatgttcttc-
accctctga gccacaacgggaactccagacttcgtcttaaatgtat
tctgacttggagagctatcacactaagcaattaacag gagctgacctggtttaggctggggtggg-
gccctactc ctcaatgttccctgaggcacatctgtgggacccctgg
gcatcatctatctgagcagccttagagctgctcatcc agttgactgttgatgtagaagtgcaaac-
ttctgcctt ccttatttgttgctttcttttttcattgttctctccc ctttgtgtctttaag
CAAGGGCATCGTAGAGCCTCCAGAAGGGACTAAGATT
TACAAGGATTCCTGGGATTTTGGCCCATATTTGAATA TCTTGAATGCTGCTATAGGAGATGA-
AATATTTCGTCA CTCATCCTGGATAAAAGAATACTTCACTTGGGCTGGA
TTTAAGGATTATAACCTGGTGGTCAGG gtatgctatgaagttattatttgtttttg- ttttcttg
tattacagagctatatgaaaacctcttagtattccag
ttggtttctcaaaagcattcattgagccttactgact gtcagacggagggcgtattggactatgt-
gctgaaaca atcctttgttgaaaatgtagggaatgttgaaaatgta
gggaatgaaatgtagatccagctctgtttctcttttg gaggattctttttcctccatcaccgtgt-
cttggttct tgtttgttttgggtttttgtgggtgttgtattgtgtt
gtgttggttatggcagtgacagctatttaaactgtga aacgggggagttcccgtcgtggcgcagt-
ggttaacga atccgactgggaaccatgaggttgcgggttcggtccc
tgcccttgctcagtgggttaacgatccggcgttgccg tgagctgtggtgtaggttgcagacacgg-
ctcggatcc cgcgttgctgtggctctagcgtaggccagcggctaca
gctccgattggacccctagcctgggaacctccatatg ccgcaggagcggcccaaagaaatagcaa-
aaagacaaa ataaataaataaataaataagtaagtaaaataaactg
tgaaacggggagttcccttcatggctcagcagttaac aaacccagctaggatccatgaggatgta-
ggttcgatc cctggccttgctcagtgggttaagaatccagcgttgc
tgtgagctgtgatgtaggtcgcagatgcagcccagat cctgcattgctgtggctgtggcgtaggc-
tggcagctg aagctccgattcaacccctagcctgggaacatccata
tgctgcaggtgtggccttaagaggcaaaaaaataaaa aaataaaaaataaataaattgtgggaca-
gacaggtgg ctccactgcagagctggtgtcctgtagcagcctggaa
gcaggtaaggtaaggactgcagctgggtaaggactga attgcaccaactgggaagtaagcctaga-
tctagaact taagttagccctgacatagacacacagagctcaccag
ctaagtggttcagcttataagctggtcactgaaactg aggatgtccacaaaagcaaaataagtag-
caacaggca gcgggatgcaagagaaagaggaggcctaaaatggtct
gggaatccctgccatacctatattttatcctacttat atttagtgcctgaatgtgtgcctggaga-
gcaaagttt agggaaagcatcgggaaatgcacagtattcataccct
taggaacaaagatcagttacctccagggtaaagacta tttccaagtttaaatttcaacccctgaa-
cattagtac tgggtaccaggcaacacttgccatcctcaaaatcaat
gaatcctaaaattcaacctgggggtcagtgacagtct gtgacaaagtttttgctggtcagtaacg-
aaataagta tgagcaccatctgagtatggtcaccaagatgtcaact
ctctttcctttggacgaattgtcattattccaagatt aggtcctttctatttttgaggtgtgaaa-
acatctttc ctttcataaaataaaaggatagtaggtggaagaattt
tttttgttttttggtctttttgctatttctttgggcc gcttctgcagcatatggaggttcccagg-
ccaggggtc gaatcggagctttagccaccggcccacgccagagcca
cagcaacacgggatccaagccgcatctgcagcctaca ccacagctcacggcaatgccggatcgtt-
aacccactg agcaagggcagggaccgaacccgcaacctcatggttc
ctagtcggattcgttaaccactgcgccacgacgggaa ctcctaatgatactcttttatatttagc-
tactatgtg atgatgagaaacagtccacattttattattttttagc
caatttgatatctcattactaagataatgataatttt ctctataaattttatttaagttagtgtt-
atgaagtgg ttttgctagtgtagaaggctaggatttgaattcagtt
caagaaagaagagagggagggagggagagggatgggt agagggatggggcagtgggagagagcaa-
agaggagag acagtttttgtattaattctgcttcattgctatcatt
taagggcacttgggtcttgcacattctagaattttct aaggaccttgaccgccagattgatatgc-
ttcttccct ttaccatgttgtcatttgaacag
ATGATTGAGACAGATGAGGACTTCAGCCCTTTGCCTG GAGGATATGACTATTTGGTTGACTTTCT-
GGATTTATC CTTTCCAAAAGAAAGACCAAGCCGGGAACATCCATAT GAGGAA
gtaagcaggaataccagtggagtgcccctttcttcct
tccttcctaaataaacttttttattttggaacaactt tagagttacagaaaagttgcaaagatat-
tatagacag tagtgtttatatatatatataaatttttttttgcttt
ttatgaccacacctgtggcatatggaggttcccagtc taggggttgaattggagctacagctgcc-
agtctgtgc cataaccacagcaatgcaggatctgggccacgtctgt
gacctacaccaaagctcacagctggattcttaaccca ctgagcaaggccagggattgaacctgca-
tcctcgtgg ttcctagttggattcgtttccgctttgccgcaatggg
aactccaaattattgttaatatcttactttactgggg tacatttgttacaaccaatactctgata-
ctgaaacat tactgttaactccgtacttgcttctttttgagtcatt
tgcaaagactggcttcttgacctgcttccttccaaac agctggcctgcctatgctgttctcagac-
ctgcaagca ctgatctctgccccccttgccttctctccagtggtgt
ctccttccccaaacaaacccagtgtggctctggaaag ggagttaagtcaacataaaccaacacat-
attttgttg agctccaattttgagcaaatccctcacctacggcaga
caggcatgatgttaagaactagggctttggacacaag gtcaagaccaagaagggttcctcacccc-
tactgattc agataaccaataatgaggctttgaatccctgtccaaa
ggttgttttttttcccttctattgagcttcttgccac cttatcagttttttttatgacagtcaaa-
tgacatgat atatgtgagcatacatggtaatttttaattctatata
aatgaatcactaaataaattaggaggatatatagtcc acctttaagcgtattacacgtgtcacat-
gaatgtgtg gcgacttaattgtagaggtttaaatgtagcttcctat
aatagatgtgttcctaaactacattttaatcattgga cttgtatttttatgttagcacttgctgt-
tgaagaaaa gcctatgccaaaagttcagtgaaaccaataatccact
gccagctttctgagttaaaaaaaatccctgggttttc acacacaggaacaccctgtgtgaaacac-
tcatttaga gcaaaatgcatctgataaggagttcctgttgtgcctc
aactggttaaggacctgacattctccatgagaatgtg agtttgatccccggccccactcgatggg-
ttaaggatc tggtgttgccacaaactgcagctccgattcatctcct
agcctagaaacttccacagcccagaatatgccacaga attcggctgtttaaaaaaaaaaagaaaa-
aaaaaagaa tcataaatgtgttggtttgttcaccaaatacatgata
acttgctcttgccaagctcagcttcataaatattaag tcatttaatacagcagccaccttatgaa-
cagatatta ctatacttcccatttacagataaggaaaatgccatat
ttaaccaagagattaaataactttcccgaggtcttat agcaagtaaatcatggtgcaggggtttg-
accacacgc agtctatctccagagtctgtgtatttagccactgttt
tactttcaaatttaaatttataaaacttctaaattat ctgttaaccataatctttggaattttta-
aaaccacga gttcctataaaatgtttcattgaaagtaagtcacttt
tccatagcttttgataatacatctgtaggataaagta agccacagctctcttgcagacttggtac-
accctgggg caaagcatcatgcctgtcacgtacatggtggtcctta
ctttgactctcagtgcttttattgcccaggaattttg tgagatttctagttgttgaggtttgttt-
aaagaggtt atgccggtacttggaagagctcttttcttgctacctg
gagccttctcatatttcctttttgaggagggacatga attgcctttcaaactcataaatatattt-
tctagtaca caagtctccatcttccttagacgcatggctcctggag
ttctccatcctcctgctccactttgggtgggctcctc tctgggtctgccaccaatctgccaccca-
gagacatcc ttgacccacttccagaccccaccatggcttcactttc
ttcgctttcctcctttgtggaaccttctgcttaagaa tctgaggaagaaaatttgcacgtgagct-
aaactggag gtacttcctgcctggtcttgcacgatagcttggctga
gcccatgatgctgggtggctgttactttccatggaca cccgaaggcgttgctcctttggcttcta-
gttgcatgc agtgttgcttatcccaggctgatctttcttccactgt
aggtgacttttaagaattaagggattaatctatatct acaacaacaacaacaaagaccttttcaa-
gctgaggta gggctttctgtatatgtttggagtggttatccagcag
actttacttgaaggcaggggtcatatcctcaagtgct cataaacggaccacagaaagatctcata-
attgggtgg agctgggtggggaccgtgtcatgtggccaggaaatgc
cagatgggaagggagtggcccttactgagctccagct gaactctgaattttctagaaaactcaga-
aatctggat ttttcatgtgtaatacccagatttatagatgtggaaa
gctaattctttttttttttaagggactataggcaatg aactaagatctaggttgtatttggacaa-
ggggtcatc agtttaagctgtgtagttgagcgctcagctattgggc
tgagggacccctaaatactgagacggggaggtccttg ctctggggcatcacaagtacactccctg-
gtctcattc aaacacttttcctacaaaattgatcccatttcttcag
tgcactgtctgaatgcatttggcccagagccgtgctg aggcatagggaaggggtccacggtttca-
tggcatcgt tttgtgctgtgtgtccctgctgtcgtccaggatacct
acctctcctcctcctgcatctgaatgtccccccacag actctctgggattctacagcctctggcc-
tgttcctca gacacctcttacctgccagctttccagattcacatta
gttagtccaaatctactgccgtcagtgactcacttca tttcttcttctccgaggcagttcagccc-
ggtacagtt gttttgtcaacacttcagttgagtctggaagatgtgc
atgggttatgcacgagagcggtccatcattttgagct agaagtcctttctcagcccagagacaag-
tcctcatct cctttacttcctgactcttcttcctctgcatccttcc
aagatatctctttctccagccaccacctaaatctctt cttttcccggggttccgtgctcaaccca-
ctcttcttc ttaaatctgtggctgggtgaacgcatctgctggcacc
acttctctgctaaagactccaaaaatccataggtcct gcccggcctttgcccacctctctccaac-
actgtccag ctttagatgtagagctaatccccccagagatatcatt
ccctggatgtctaagtcctttggtatctcactttcag cgtgttcaaaatcctcttacaactgttc-
tttctcctt ttccatcttgattattggcaacatgccagcctttccc
ctacccccagcagtgagccaagctagaaacaagggct taatcttcaatctttccttctccatccc-
taaacctaa tgagtctccaagcccttcccagtttacaccctaaatg
ttgctcaaaacatcccctagttcttccacgtgctctc ctctatattgaaaggtcaagaaaggcca-
tcttccctc cactgtgaggaaatagatcttgatactgcccctgagc
tgggcagtcctcgacctgacaaactgtgcagtgtttc taaatctctactggcaaaatgagagtgc-
ctttgacct gtgttgcgatctcagatcacagtggatgtaattgttt
tataggaatggtgaacgaaaaagaagtaaatccctaa tgccaaactcctgatcattctatgtcat-
ttaatagcc tgtcatttatgataaagtttcctctactggcattagc
acaatacttctcaggaaaaaaaaatatgatgccagat actgaaaagctcctgggtaaacatgaac-
atgggtacc gataaaatggtgaagccagtccaatcttagagtgact
tcccttcatgctacttcatgctctttttttttttttt ttttaagaaaaaccccttttttttttct-
cacaccagt cacagaggagaccgaggcttagcaaggttaaggtcac
atgattagtaagtgctgggctgaaactcaaaaccatc tctgcttgtctcctaaccctgtgcacct-
ctgactatt caacag ATCCTGTGTCAGGAGTTGGGATTCTTTGAAG
gtaagggccttgaccaccgaattaaggtaatcttgct
ctgtggcaggccttgttttcagtattttaagtacact ggctcaggtaatcctcacaacagcccca-
ggaggaatg ttctattacctccactgtatagatgaggaacttgagg
cacagaatggttgccaaggtcacacagctatattggg ggttcatacccagccatccaactctgtc-
tgtactctc tgccactctgcacccccagctcctgatccacttcctg
tttccatccctcgatttctgctgcactcaggggcccc tctccccctcggcctgtgagatctgctt-
cagtaggct tttctccctgactcctccatccctgtccttacaggca
gctgcttctctccgggacacgaggggtccatacggac actctctactggctgggttgcgcctaac-
tcgtgattc ctcctctgtttcag ATTCGGAGCCGGGTTGATGTCATCAG- ACACGTGGTAA
AGAATGGTCTGCTCTGGGATGACTTGTACATAGGATT
CCAAACCCGGCTTCAGCGGGATCCTGATATATACCAT CATCT
gtaagtccgaaaatgcctgtcgtgtgtgccttaggct gctgcggaggaggccagggctatataag-
cagagtcag tgactgactgtgccctgcagtgttgatggccatggag
attccaccgttagagcttttttctttgttaaccttga aggcaaatctggttaggaagataacttt-
caaagagtc accatctggacattcatgcccatgtgcttcaatcctg
tatacaagcagtttagagtacagggaagggaaggaca ttatgaaagggagagggtgtgtttggat-
ccagcagct ccatcctcagaatttatctgaagacactgcaaaatta
ctaagaatcactatgacaagaatgaggatggggtgat atggcaaagttgtgatcctggaagacct-
tcatctccc atgttgcccaactctgaacatgaatttggtgaactag
ttggttaaggggatgatcctccaagtttctccctggt tgagctccaaaaaccatgtaagtttctc-
atagcaaaa ccgtataggtccttagggctttagttggaatatttgt
gctgaaatgctggaaagccccatttgccatttttgta tttgcaaaataatcatcaagaggggaga-
atgcattct ttcatgaccactgaccctctgaaaaggtcaggaattt
agtctgaagtaggcaagcctcctaccccgcttctgcc atgagcttgcacgcacaggcctgtcttg-
acatttctt ctttatagatttctttttgaatatcttgaaattgctt
taaaaatatttaaagaatgtagaattatataaaataa aaaggaaataaccccacacctcccacaa-
aaccctgtt tcctgcctttctccacccactetccagggtaacactt
ggtaacagcatagttgtatcaccccaggcctattttt gagcatatcagcatttcaagaaatgtat-
tttttctca ataaaacatcccttatagttgaggaggggaggttatc
attcctgggttttgttttttttttttttttaatgtaa tcctggtacatcggtaatttgcattttt-
tattcatta atatctttggtatttctagtgttgggacacacaggtc
aacctcagtttttgggtttttttttttgtctttttgt ctttctagggccacacctgcagcatatg-
gacgttccc aagctaggagtctaatcagagctgtagccaccagcct
acgtcatagccatagcaacgtcagatccaagccgtgt ctgtgacctacaagcacagctcatggca-
acaccggat ccttaaccactgaacgaggccaggggatcgaacacac
atcctcatggatcctagtcatgttcattaaccactga gtcatgatgggaactccaacttcaacta-
ttttaatgt ctgtaaaacattccatttggaaaccatttcatttgta
aagcaaaatgaaaacattttgttcattttcaacagag ttcgtagctgacttctgttctggaaaaa-
aggaaatgg agcaaatttgagtgagaaagattcaaagataactttt
cttttaaaaaaaattatatcttggaaacttctgggct attgattctgaagactatttttctatat-
actgttttg atagcaaagttcataaatgtgaaaggatcctgcgatg
aatcttgggaagcagtcatagcccaatatatctttgt tgcttttaaaatgagatttagtttacta-
aatattttt ctgatcataaaaataacacagatctaccgcagaaaat
ttggaaaaaaaaaaacttttaaattcaaaaaacagtt aaaccacaaatgatcccaccatccagag-
agcaatttg tactttggtgtctagttcatctttctttttctgttta
caagcacatataccacaagcattttttcaaaaaatga aaatgggataatactatacatacgtctg-
tacacctgc atagttactgaacagtctttgatctaccctgtaagtt
tctaacttttcattatttgaaatgatgttttggcaaa gaaatatgtaggtgtgtctcgcacactt-
tcataatga tttcttaggataaatttcttaggataaattcataatg
atttcttataataatccatactctgccaactgatctt cagggaagccaactcgccttctcagaaa-
taacatata acccatttacttgccctctcaccaatactaggtccta
atgtttttgtgtacagattctatatttttacatacaa gaattccttaaagcaaggcatgtcacag-
aaaaataga aggaagacacaattgtcatgtttaaggactgcattct
gtaccaaaaatgctaagttaaatgaacatctgaaaca gtacagaaacgctatctttcagggaaag-
ctgagtacc aggtactgaacagattttggcaaatacagcaggcatg
gatgtttccaaaacatgtttttctactttatctctta cag
GTTTTGGAATCATTTTCAAATAAAACTCCCCCTCACA CCACCTGACTGGAAGTCCTTCCTGATGT-
GCTCTGGGT AGAGAGGACCTGAGCTGTCCCAG
gtaaagcatcctgcaggtctgggagacactcttattc tccagcccatcacactgtgtttggcatc-
agaattaag caggcactatgcctatcagaaaacctgacttttgggg
gaatgaaagaagctaacattacaagaatgtctgtgtt taaaaataagtcaataagggagttccca-
tcgtggctc agtggtaacgaaccctactagtatccattgaggacac
aggttcaatatctggcctcactcagtcggctaaggat ccagtgatgccgtgagctgcagtgtagg-
ccacagacg tggctcagatctggtgctgctgtggctatggtgtagg
ccggccccctgtaactccaattcgacccctaggctgg gaacctaaaaagaccccaaaaaagtcgc-
tttaatgaa tagtgaatacatccagcccaaagtccacagactcttt
ggtctggttgtggcaaacatacagccagttaacaaac aagacaaaaattatcctaggtggtcagt-
gggggttca gagctgaatcctgaacactggaaggaaaacagcaacc
aaatccaaatactgtatggttttgcttatatgtagaa tctaaattcaaagcaaatgagcaaacca-
attgaaaca gttatggaagacaagcaggtggttgtcaggggggaga
taaggggaggcaggaaagacctgggcgagggagatta agaggtaccaactttcagttgcaaaaca-
aatgagtca ccagtatgaaatgtgcaatgtgggaaatacaggccat
aactttataatctcttttttttttttgtcttttttgc cttttctaaggctgctcccgtggcatat-
ggaggttcc caggctaggagtccaaacagagctgtagctgccagcc
tacaccagagccacagcaacacgggaaccttaacccg ctgagcaaggccagggatcgaacccgag-
tcctcacag atgccagtagggttcattaaccactgagccacgacag
gaattccagggtctgttgtgttcttaaaacacttcca ggagagtgagtggtatgtcataagtaaa-
caataaatg ttaaccacaacaagcttatgaaataaacaggaaagcc
atatgacctacaatcagtcattgggagaatccacaaa aggttgagcagaggatcaattccagctc-
acactccag ttttagattctcccctgccttaaagcatcacagacta
cataatctgagctgaagaataaaaattaaaactcacc ccagtgcaaaacagaaatgaaaaagtat-
taaaacgag gttcatactgttgttcattagcaatatcttttattca cag
GGGTGCCCAACAACATGAAAAAATCAAGAATTTATTG
CTGCTACGTCAAAGCTTATACCAGAGATTATGCCTTA TAGACATTAGCAATGGATAATTATATGT-
TGCACTTGT GAAATGTGCACATATCCTGTTTATGAATCACCACATA
GCCAGATTATCAATATTTTACTTATTTCGTAAAAAAT CCACAATTTTCCATAACAGAATCAACGT-
GTGCAATAG GAACAAGATTGCTATGGAAAACGAGGGTAACAGGAGG
AGATATTAATCCAAGCATAGAAGAAATAGACAAATGA GGGGCCATAAGGGGAATATAGGGAAGAG-
AAAAAAATT AAGATGGAATTTTAAAAGGAGAATGTAAAAAATAGAT
ATTTGTTCCTTAATAGGTTGATTCCTCAAATAGAGCC CATGAATATAATCAAATAGGAAGGGTTC-
ATGACTGTT TTCAATTTTTCAAAAAGCTTTGTTGAAATCATAGACT
TGCAAAACAAGGCTGTAGAGGCCACCCTAAAATGGAA AATTTCACTGGGACTGAAATTATTTTGA-
TTCAATGAC AAAATTTGTTATTTACTGCGGATTATAAACTCTAACA
AATAGCGATCTCTTTGCTTCATAAAAACATAAACACT
AGCTAGTAATAAAATGAGTTCTGCAG
[0094] SEQ ID NO. 50 represents contiguous genomic sequences
containing Exon 12, Intron 12, Exon 13, Intron 13, Exon 14, Intron
14, Exon 15, Intron 15, Exon 16, Intron 16, Exon 17, Intron 17, and
Exon 18. Nucleotide sequences that contain at least 150, 200, 250,
300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900,
950, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500,
6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, 11000,
12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000 or 20,000
contiguous nucleotides of SEQ ID NO. 50 are provided, as well as
nucleotide sequences at least 80, 85, 90, 95, 98, or 99% homologous
to SEQ ID NO. 50.
VIII. Oligonucleotide Probes and Primers
[0095] The present invention further provides oligonucleotide
probes and primers which hybridize to the hereinabove-described
sequences (SEQ ID Nos. 1, 3, 5, 7, 9-45, 46, 47, 48, 49 and 50).
Oligonucleotides are provided that can be homologous to SEQ ID Nos.
1, 3, 5, 7, 9-45, 46, 47, 48, 49 and 50, and fragments thereof.
Oligonucleotides that hybridize under stringent conditions to SEQ
ID Nos. 1, 3, 5, 7, 9-45, 46, 47, 48, 49 and 50 and fragments
thereof, are also provided. Stringent conditions describe
conditions under which hybridization will occur only if there is at
least about 85%, about 90%, about 95%, or at least about 98%
homology between the sequences. Alternatively, the oligonucleotide
can have at least 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,
32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 45, 46, 47, 48, 49,
50, 75 or 100 bases which hybridize to SEQ ID Nos. 1, 3, 5, 7,
9-45, 46, 47, 48, 49 and 50, and fragments thereof. Such
oligonucleotides can be used as primers and probes to detect the
sequences provided herein. The probe or primer can be at least 14
nucleotides in length, and in a preferred embodiment, are at least
15, 20, 25, 28, 30, or 35 nucleotides in length.
[0096] Given the above sequences, one of ordinary skill in the art
using standard algorithms can construct oligonucleotide probes and
primes that are complementary to sequences contained in Seq ID Nos.
1, 3, 5, 7, 9-45, 46, 47, 48, 49 and 50, and fragments thereof. The
rules for complementary pairing are well known: cytosine ("C")
always pairs with guanine ("G") and thymine ("T") or uracil ("U")
always pairs with adenine ("A"). It is recognized that it is not
necessary for the primer or probe to be 100% complementary to the
target nucleic acid sequence, as long as the primer or probe
sufficiently hybridizes and can recognize the corresponding
complementary sequence. A certain degree of pair mismatch can
generally be tolerated.
[0097] Oligonucleotide sequences used as the hybridizing region of
a primer can also be used as the hydridizing region of a probe.
Suitability of a primer sequence for use as a probe depends on the
hybridization characteristics of the primer. Similarly, an
oligonucleotide used as a probe can be used as a primer.
[0098] It will be apparent to those skilled in the art that,
provided with these specific embodiments, specific primers and
probes can be prepared by, for example, the addition of nucleotides
to either the 5' or 3' ends, which nucleotides are complementary to
the target sequence or are not complimentary to the target
sequence. So long as primer compositions serve as a point of
initiation for extension on the target sequences, and so long as
the primers and probes comprise at least 14 consecutive nucleotides
contained within the above mentioned SEQ ID Nos. such compositions
are within the scope of the invention.
[0099] The probes and primers herein can be selected by the
following criteria, which are factors to be considered, but are not
exclusive or determinative. The probes and primers are selected
from the region of the CMP-Neu5Ac hydroxylase nucleic acid sequence
identified in SEQ ID Nos. 1, 3, 5, 7, 945, 46, 47, 48, 49, 50, and
fragments thereof. The probes and primers lack homology with
sequences of other genes that would be expected to compromise the
test. The probes or primers lack secondary structure formation in
the amplified nucleic acid which can interfere with extension by
the amplification enzyme such as E. coli DNA polymerase, preferably
that portion of the DNA polymerase referred to as the Klenow
fragment. This can be accomplished by employing up to about 15% by
weight, preferably 5-10% by weight, dimethyl sulfoxide (DMSO) in
the amplification medium and/or increasing the amplification
temperatures to 30.degree.-40.degree. C.
[0100] Preferably, the probes or primers should contain
approximately 50% guanine and cytosine nucleotides, as measured by
the formula adenine (A)+thymine (T)+cytosine (C)+guanine
(G)/cytosine (C)+guanine (G). Preferably, the probe or primer does
not contain multiple consecutive adenine and thymine residues at
the 3' end of the primer which can result in less stable
hybrids.
[0101] The probes and primers of the invention can be about 10 to
30 nucleotides long, preferably at least 10, 11, 12, 13, 14, 15,
20, 25, or 28 nucleotides in length, including specifically 14, 15,
16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30
nucleotides. The nucleotides as used in the present invention can
be ribonucleotides, deoxyribonucleotides and modified nucleotides
such as inosine or nucleotides containing modified groups which do
not essentially alter their hybridization characteristics. Probe
and primer sequences are represented throughout the specification
as single stranded DNA oligonucleotides from the 5' to the 3' end.
Any of the probes can be used as such, or in their complementary
form, or in their RNA form (wherein T is replaced by U).
[0102] The probes and primers according to the invention can be
prepared by cloning of recombinant plasmids containing inserts
including the corresponding nucleotide sequences, optionally by
cleaving the latter out from the cloned plasmids upon using the
adequate nucleases and recovering them, e.g. by fractionation
according to molecular weight. The probes and primers according to
the present invention can also be synthesized chemically, for
instance by the conventional phosphotriester or phosphodiester
methods or automated embodiments thereof. In one such automated
embodiment diethylphosphoramidites are used as starting materials
and can be synthesized as described by Beaucage, et al.,
Tetrahedron Letters 22:1859-1862 (1981). One method of synthesizing
oligonucleotides on a modified solid support is described in U.S.
Pat. No. 4,458,066. It is also possible to use a probe or primer
which has been isolated from a biological source (such as a
restriction endonuclease digest).
[0103] The oligonucleotides used as primers or probes can also
comprise nucleotide analogues such as phosphorothiates (Matsukura
S., Naibunpi Gakkai Zasshi. 43(6):527-32 (1967)),
alkylphosphorothiates (Miller P., et al., Biochemistry
18(23):5134-43 (1979), peptide nucleic acids (Nielsen P., et al.,
Science 254(5037):1497-500 (1991); Nielsen P., et al.,
Nucleic-Acids-Res. 21(2):197-200 (1993)), morpholino nucleic acids,
locked nucleic acids, pseudocyclic oligonucleobases,
2'-O,4'-C-ethylene bridged nucleic acids or can contain
intercalating agents (Asseline J., et al., Proc. Natl. Acad. Sci.
USA 81(11):3297-301 (1984)).
[0104] For designing probes and primers with desired
characteristics, the following useful guidelines known to the
person skilled in the art can be applied. Because the extent and
specificity of hybridization reactions are affected by a number of
factors, manipulation of one or more of those factors will
determine the exact sensitivity and specificity of a particular
probe, whether perfectly complementary to its target or not. The
importance and effect of various assay conditions, explained
further herein, are known to those skilled in the art.
[0105] The stability of the probe and primer to target nucleic acid
hybrid should be chosen to be compatible with the assay conditions.
This can be accomplished by avoiding long AT-rich sequences, by
terminating the hybrids with GC base pairs, and/or by designing the
probe with an appropriate Tm. The beginning and end points of the
probe should be chosen so that the length and % GC result in a Tm
about 2-10.degree. C. higher than the temperature at which the
final assay will be performed. The base composition of the probe is
significant because G-C base pairs exhibit greater thermal
stability compared to A-T base pairs due to additional hydrogen
bonding. Thus, hybridization involving complementary nucleic acids
of higher G-C content will be stable at higher temperatures.
Conditions such as ionic strength and incubation temperature under
which probe will be used should also be taken into account when
designing a probe. It is known that hybridization will increase as
the ionic strength of the reaction mixture increases, and that the
thermal stability of the hybrids will increase with increasing
ionic strength. Chemical reagents, such as formamide, urea, DIVISO
and alcohols, which disrupt hydrogen bonds, will increase the
stringency of hybridization. Destabilization of the hydrogen bonds
by such reagents can greatly reduce the Tm. In general, optimal
hybridization for synthetic oligonucleotide probes of about 10-50
bases in length occurs approximately 5.degree. C. below the melting
temperature for a given duplex. Incubation at temperatures below
the optimum can allow mismatched base sequences to hybridize and
can therefore result in reduced specificity. It is desirable to
have probes which hybridize only under conditions of high
stringency. Under high stringency conditions only highly
complementary nucleic acid hybrids will form; hybrids without a
sufficient degree of complementarity will not form. Accordingly,
the stringency of the assay conditions determines the amount of
complementarity needed between two nucleic acid strands forming a
hybrid. The degree of stringency is chosen such as to maximize the
difference in stability between the hybrid formed with the target
and the non-target nucleic acid. In the present case, single base
pair changes need to be detected, which requires conditions of very
high stringency.
[0106] The length of the target nucleic acid sequence and,
accordingly, the length of the probe sequence can also be
important. In some cases, there can be several sequences from a
particular region, varying in location and length, which will yield
probes and primers with the desired hybridization characteristics.
In other cases, one sequence can be significantly better than
another which differs merely by a single base.
[0107] While it is possible for nucleic acids that are not
perfectly complementary to hybridize, the longest stretch of
perfectly complementary base sequence will normally primarily
determine hybrid stability. While oligonucleotide probes and
primers of different lengths and base composition can be used,
preferred oligonucleotide probes and primers of this invention are
between about 14 and 30 bases in length and have a sufficient
stretch in the sequence which is perfectly complementary to the
target nucleic acid sequence.
[0108] Regions in the target DNA or RNA which are known to form
strong internal structures inhibitory to hybridization are less
preferred. Likewise, probes with extensive self-complementarity
should be avoided. As explained above, hybridization is the
association of two single strands of complementary nucleic acids to
form a hydrogen bonded double strand. It is implicit that if one of
the two strands is wholly or partially involved in a hybrid, it
will be less able to participate in formation of a new hybrid.
There can be intramolecular and intermolecular hybrids formed
within the molecules of one type of probe if there is sufficient
self complementarity. Such structures can be avoided through
careful probe design. By designing a probe so that a substantial
portion of the sequence of interest is single stranded, the rate
and extent of hybridization can be greatly increased. Computer
programs are available to search for this type of interaction.
However, in certain instances, it may not be possible to avoid this
type of interaction.
[0109] Specific primers and sequence specific oligonucleotide
probes can be used in a polymerase chain reaction that enables
amplification and detection of CMP-Neu5Ac hydroxylase nucleic acid
sequences.
IV. Genetic Targeting of the CMP-Neu5Ac Hydroxylase Gene
[0110] Gene targeting allows for the selective manipulation of
animal cell genomes. Using this technique, a particular DNA
sequence can be targeted and modified in a site-specific and
precise manner. Different types of DNA sequences can be targeted
for modification, including regulatory regions, coding regions and
regions of DNA between genes. Examples of regulatory regions
include: promoter regions, enhancer regions, terminator regions and
introns. By modifying these regulatory regions, the timing and
level of expression of a gene can be altered. Coding regions can be
modified to alter, enhance or eliminate the protein within a cell.
Introns and exons, as well as inter-genic regions, are suitable
targets for modification.
[0111] Modifications of DNA sequences can be of several types,
including insertions, deletions, substitutions, or any combination
thereof. A specific example of a modification is the inactivation
of a gene by site-specific integration of a nucleotide sequence
that disrupts expression of the gene product, i.e. a "knock out".
For example, one approach to disrupting the CMP-Neu5Ac hydroxylase
gene is to insert a selectable marker into the targeting DNA such
that homologous recombination between the targeting DNA and the
target DNA can result in insertion of the selectable marker into
the coding region of the target gene. For example, see FIGS. 3, 12,
and 13. In this way, for example, the CMP-Neu5Ac hydoxylase gene
sequence is disrupted, rendering the encoded enzyme
nonfunctional.
Homologous Recombination
[0112] Homologous recombination permits site-specific modifications
in endogenous genes and thus novel alterations can be engineered
into the genome. A primary step in homologous recombination is DNA
strand exchange, which involves a pairing of a DNA duplex with at
least one DNA strand containing a complementary sequence to form an
intermediate recombination structure containing heteroduplex DNA
(see, for example Radding, C. M. (1982) Ann. Rev. Genet. 16: 405;
U.S. Pat. No. 4,888,274). The heteroduplex DNA can take several
forms, including a three DNA strand containing triplex form wherein
a single complementary strand invades the DNA duplex (Hsieh, et
al., Genes and Development 4: 1951 (1990); Rao, et al., (1991) PNAS
88:2984)) and, when two complementary DNA strands pair with a DNA
duplex, a classical Holliday recombination joint or chi structure
(Holliday, R., Genet. Res. 5: 282 (1964)) can form, or a double-D
loop ("Diagnostic Applications of Double-D Loop Formation" U.S.
Ser. No. 07/755,462, filed Sep. 4, 1991). Once formed, a
heteroduplex structure can be resolved by strand breakage and
exchange, so that all or a portion of an invading DNA strand is
spliced into a recipient DNA duplex, adding or replacing a segment
of the recipient DNA duplex. Alternatively, a heteroduplex
structure can result in gene conversion, wherein a sequence of an
invading strand is transferred to a recipient DNA duplex by repair
of mismatched bases using the invading strand as a template (Genes,
3.sup.rd Ed. (1987) Lewin, B., John Wiley, New York, N.Y.; Lopez,
et al., Nucleic Acids Res. 15: 5643(1987)). Whether by the
mechanism of breakage and rejoining or by the mechanism(s) of gene
conversion, formation of heteroduplex DNA at homologously paired
joints can serve to transfer genetic sequence information from one
DNA molecule to another.
[0113] The ability of homologous recombination (gene conversion and
classical strand breakage/rejoining) to transfer genetic sequence
information between DNA molecules renders targeted homologous
recombination a powerful method in genetic engineering and gene
manipulation.
[0114] In homologous recombination, the incoming DNA interacts with
and integrates into a site in the genome that contains a
substantially homologous DNA sequence. In non-homologous ("random"
or "illicit") integration, the incoming DNA is not found at a
homologous sequence in the genome but integrates elsewhere, at one
of a large number of potential locations. In general, studies with
higher eukaryotic cells have revealed that the frequency of
homologous recombination is far less than the frequency of random
integration. The ratio of these frequencies has direct implications
for "gene targeting" which depends on integration via homologous
recombination (i.e. recombination between the exogenous "targeting
DNA" and the corresponding "target DNA" in the genome).
[0115] A number of papers describe the use of homologous
recombination in mammalian cells. Illustrative of these papers are
Kucherlapati, et al., Proc. Natl. Acad. Sci. (USA) 81:3153-3157,
1984; Kucherlapati, et al., Mol. Cell. Bio. 5:714-720, 1985;
Smithies, et al, Nature 317:230-234, 1985; Wake, et al., Mol. Cell.
Bio. 8:2080-2089, 1985; Ayares, et al., Genetics 111:375-388, 1985;
Ayares, et al., Mol. Cell. Bio. 7:1656-1662, 1986; Song, et al.,
Proc. Natl. Acad. Sci USA 84:6820-6824, 1987; Thomas, et al. Cell
44:419-428, 1986; Thomas and Capecchi, Cell 51: 503-512, 1987;
Nandi, et al., Proc. Natl. Acad. Sci. USA 85:3845-3849, 1988; and
Mansour, et al., Nature 336:348-352, 1988; Evans and Kaufman,
Nature 294:146-154, 1981; Doetschman, et al., Nature 330:576-578,
1987; Thoma and Capecchi, Cell 51:503-512,4987; Thompson, et al.,
Cell 56:316-321, 1989.
[0116] The present invention uses homologous recombination to
inactivate the porcine CMP-Neu5Ac hydroxylase gene in cells, such
as fibroblasts. The DNA can comprise at least a portion of the
gene(s) at the particular locus with introduction of an alteration
into at least one, optionally both copies, of the native gene(s),
so as to prevent expression of a functional enzyme and production
of a Hanganutziu-Deicher antigen molecule. The alteration can be an
insertion, deletion, replacement or combination thereof. When the
alteration is introduce into only one copy of the gene being
inactivated, the cells having a single unmutated copy of the target
gene are amplified and can be subjected to a second targeting step,
where the alteration can be the same or different from the first
alteration, usually different, and where a deletion, or replacement
is involved, can be overlapping at least a portion of the
alteration originally introduced. In this second targeting step, a
targeting vector with the same arms of homology, but containing a
different mammalian selectable markers can be used. The resulting
transformants are screened for the absence of a functional target
antigen and the DNA of the cell can be further screened to ensure
the absence of a wild-type target gene. Alternatively, homozygosity
as to a phenotype can be achieved by breeding hosts heterozygous
for the mutation.
[0117] Porcine cells that can be genetically modified can be
obtained from a variety of different organs and tissues such as,
but not limited to, brain, heart, lungs, glands, brain, eye,
stomach, spleen, pancreas, kidneys, liver, intestines, uterus,
bladder, skin, hair, nails, ears, nose, mouth, lips, gums, teeth,
tongue, salivary glands, tonsils, pharynx, esophagus, large
intestine, small intestine, rectum, anus, pylorus, thyroid gland,
thymus gland, suprarenal capsule, bones, cartilage, tendons,
ligaments, skeletal muscles, smooth muscles, blood vessels, blood,
spinal cord, trachea, ureters, urethra, hypothalamus, pituitary,
adrenal glands, ovaries, oviducts, uterus, vagina, mammary glands,
testes, seminal vesicles, penis, lymph, lymph nodes and lymph
vessels. In one embodiment of the invention, porcine cells can be
selected from the group consisting of, but not limited to,
epithelial cells, fibroblast cells, neural cells, keratinocytes,
hematopoietic cells, melanocytes, chondrocytes, lymphocytes (B and
T), macrophages, monocytes, mononuclear cells, cardiac muscle
cells, other muscle cells, .quadrature.hosphate cells, cumulus
cells, epidermal cells, endothelial cells, Islets of Langerhans
cells, blood cells, blood precursor cells, bone cells, bone
precursor cells, neuronal stem cells, primordial stem cells,
hepatocytes, keratinocytes, umbilical vein endothelial cells,
aortic endothelial cells, microvascular endothelial cells,
fibroblasts, liver stellate cells, aortic smooth muscle cells,
cardiac myocytes, neurons, Kupffer cells, smooth muscle cells,
Schwann cells, and epithelial cells, erythrocytes, platelets,
neutrophils, lymphocytes, monocytes, eosinophils, basophils,
adipocytes, chondrocytes, pancreatic islet cells, thyroid cells,
parathyroid cells, parotid cells, tumor cells, glial cells,
astrocytes, red blood cells, white blood cells, macrophages,
epithelial cells, somatic cells, pituitary cells, adrenal cells,
hair cells, bladder cells, kidney cells, retinal cells, rod cells,
cone cells, heart cells, pacemaker cells, spleen cells, antigen
presenting cells, memory cells, T cells, B cells, plasma cells,
muscle cells, ovarian cells, uterine cells, prostate cells, vaginal
epithelial cells, sperm cells, testicular cells, germ cells, egg
cells, leydig cells, peritubular cells, sertoli cells, lutein
cells, cervical cells, endometrial cells, mammary cells, follicle
cells, mucous cells, ciliated cells, nonkeratinized epithelial
cells, keratinized epithelial cells, lung cells, goblet cells,
columnar epithelial cells, squamous epithelial cells, osteocytes,
osteoblasts, and osteoclasts.
[0118] In one alternative embodiment, embryonic stem cells can be
used. An embryonic stem cell line can be employed or embryonic stem
cells can be obtained freshly from a host, such as a porcine
animal. The cells can be grown on an appropriate fibroblast-feeder
layer or grown in the presence of leukemia inhibiting factor (LIF).
In a preferred embodiment, the porcine cells can be fibroblasts; in
one specific embodiment, the porcine cells can be fetal
fibroblasts. Fibroblast cells are a preferred somatic cell type
because they can be obtained from developing fetuses and adult
animals in large quantities.
[0119] These cells can be easily propagated in vitro with a rapid
doubling time and can be clonally propagated for use in gene
targeting procedures.
Targeting Vectors
[0120] Cells homozygous at a targeted locus can be produced by
introducing DNA into the cells, where the DNA has homology to the
target locus and includes a marker gene, allowing for selection of
cells comprising the integrated construct. The homologous DNA in
the target vector will recombine with the chromosomal DNA at the
target locus (see, for example, FIGS. 3, 12, and 13). The marker
gene can be flanked on both sides by homologous DNA sequences, a
3'recombination arm and a 5' recombination arm (See, for example,
FIG. 11). Methods for the construction of targeting vectors have
been described in the art, see, for example, Dai et al., Nature
Biotechnology 20: 251-255, 2002; WO 00/51424.
[0121] Various constructs can be prepared for homologous
recombination at a target locus. Usually, the construct can include
at least 50 bp, 100 bp, 500 bp, 1 kbp, 2 kbp, 4 kbp, 5 kbp, 10 kbp,
15 kbp, 20 kbp, or 50 kbp of sequence homologous with the target
locus. The sequence can include any contiguous sequence of the
porcine CMP-Neu5Ac hydroxylase gene, including at least 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110,
120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300,
325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 700, 750, 800,
850, 900, 1000, 5000 or 10,000 contiguous nucleotides of Seq ID Nos
9-45, 46, 47, 48, 49, and 50, or any combination or fragment
thereof. Fragments of Seq ID Nos. 9-45, 46, 47, 48, 49 and 50 can
include any contiguous nucleic acid or peptide sequence that
includes at least about 10 bp, 15 bp, 17 bp, 20 bp, 50 bp, 100 bp,
500 bp, 1 kbp, 5 kbp or 10 kpb.
[0122] Various considerations can be involved in determining the
extent of homology of target DNA sequences, such as, for example,
the size of the target locus, availability of sequences, relative
efficiency of double cross-over events at the target locus and the
similarity of the target sequence with other sequences.
[0123] The targeting DNA can include a sequence in which DNA
substantially isogenic flanks the desired sequence modifications
with a corresponding target sequence in the genome to be modified.
The substantially isogenic sequence can be at least about 95%,
97-98%, 99.0-99.5%, 99.6-99.9%, or 100% identical to the
corresponding target sequence (except for the desired sequence
modifications). The targeting DNA and the target DNA preferably can
share stretches of DNA at least about 75, 150 or 500 base pairs
that are 100% identical. Accordingly, targeting DNA can be derived
from cells closely related to the cell line being targeted; or the
targeting DNA can be derived from cells of the same cell line or
animal as the cells being targeted.
[0124] The DNA constructs can be designed to modify the endogenous,
target CMP-Neu5Ac hydroxylase. The homologous sequence for
targeting the construct can have one or more deletions, insertions,
substitutions or combinations thereof designed to disrupt the
function of the resultant gene product. In one embodiment, the
alteration can be the insertion of a selectable marker gene fused
in reading frame with the upstream sequence of the target gene.
[0125] Suitable selectable marker genes include, but are not
limited to: genes conferring the ability to grow on certain media
substrates, such as the tk gene (thymidine kinase) or the hprt gene
(hypoxanthine phosphoribosyltransferase) which confer the ability
to grow on HAT medium (hypoxanthine, aminopterin and thymidine);
the bacterial gpt gene (guanine/xanthine phosphoribosyltransferase)
which allows growth on MAX medium (mycophenolic acid, adenine, and
xanthine). See, for example, Song, K-Y., et al. Proc. Nat'l Acad.
Sci. U.S.A. 84:6820-6824 (1987); Sambrook, J., et al., Molecular
Cloning--A Laboratory Manual, Cold Spring Harbor Laboratory, Cold
Spring Harbor, N.Y. (1989), Chapter 16. Other examples of
selectable markers include: genes conferring resistance to
compounds such as antibiotics, genes conferring the ability to grow
on selected substrates, genes encoding proteins that produce
detectable signals such as luminescence, such as green fluorescent
protein, enhanced green fluorescent protein (eGFP). A wide variety
of such markers are known and available, including, for example,
antibiotic resistance genes such as the neomycin resistance gene
(neo) (Southern, P., and P. Berg, J. Mol. Appl. Genet. 1:327-341
(1982)); and the hygromycin resistance gene (hyg) (Nucleic Acids
Research 11:6895-6911 (1983), and Te Riele, H., et al., Nature
348:649-651 (1990)). Other selectable marker genes include:
acetohydroxy acid synthase (AHAS), alkaline phosphatase (AP), beta
galactosidase (LacZ), beta glucoronidase (GUS), chloramphenicol
acetyltransferase (CAT), green fluorescent protein (GFP), red
fluorescent protein (RFP), yellow fluorescent protein (YFP), cyan
fluorescent protein (CFP), horseradish peroxidase (HRP), luciferase
(Luc), nopaline synthase (NOS), octopine synthase (OCS), and
derivatives thereof. Multiple selectable markers are available that
confer resistance to ampicillin, bleomycin, chloramphenicol,
gentamycin, hygromycin, kanamycin, lincomycin, methotrexate,
phosphinothricin, puromycin, and tetracycline.
[0126] Methods for the incorporation of antibiotic resistance genes
and negative selection factors will be familiar to those of
ordinary skill in the art (see, e.g., WO 99/15650; U.S. Pat. No.
6,080,576; U.S. Pat. No. 6,136,566; Niwa, et al., J. Biochem.
113:343-349 (1993); and Yoshida, et al., Transgenic Research,
4:277-287 (1995)).
[0127] Additional selectable marker genes useful in this invention,
for example, are described in U.S. Pat. Nos. 6,319,669; 6,316,181;
6,303,373; 6,291,177; 6,284,519; 6,284,496; 6,280,934; 6,274,354;
6,270,958; 6,268,201; 6,265,548; 6,261,760; 6,255,558; 6,255,071;
6,251,677; 6,251,602; 6,251,582; 6,251,384; 6,248,558; 6,248,550;
6,248,543; 6,232,107; 6,228,639; 6,225,082; 6,221,612; 6,218,185;
6,214,567; 6,214,563; 6,210,922; 6,210,910; 6,203,986; 6,197,928;
6,180,343; 6,172,188; 6,153,409; 6,150,176; 6,146,826; 6,140,132;
6,136,539; 6,136,538; 6,133,429; 6,130,313; 6,124,128; 6,110,711;
6,096,865; 6,096,717; 6,093,808; 6,090,919; 6,083,690; 6,077,707;
6,066,476; 6,060,247; 6,054,321; 6,037,133; 6,027,881; 6,025,192;
6,020,192; 6,013,447; 6,001,557; 5,994,077; 5,994,071; 5,993,778;
5,989,808; 5,985,577; 5,968,773; 5,968,738; 5,958,713; 5,952,236;
5,948,889; 5,948,681; 5,942,387; 5,932,435; 5,922,576; 5,919,445;
and 5,914,233.
[0128] Combinations of selectable markers can also be used. For
example, to target CMP-NeuSAc hydroxylase, a neo gene (with or
without its own promoter, as discussed above) can be cloned into a
DNA sequence which is homologous to the CMP-Neu5Ac hydroxylase
gene. To use a combination of markers, the HSV-tk gene can be
cloned such that it is outside of the targeting DNA (another
selectable marker could be placed on the opposite flank, if
desired). After introducing the DNA construct into the cells to be
targeted, the cells can be selected on the appropriate antibiotics.
In this particular example, those cells which are resistant to G418
and gancyclovir are most likely to have arisen by homologous
recombination in which the neo gene has been recombined into the
CMP-Neu5Ac hydroxylase gene but the tk gene has been lost because
it was located outside the region of the double crossover.
[0129] Deletions can be at least about 50 bp, more usually at least
about 100 bp, and generally not more than about 20 kbp, where the
deletion can normally include at least a portion of the coding
region including a portion of or one or more exons, a portion of or
one or more introns, and can or can not include a portion of the
flanking non-coding regions, particularly the 5'-non-coding region
(transcriptional regulatory region). Thus, the homologous region
can extend beyond the coding region into the 5'-non-coding region
or alternatively into the 3'-non-coding region. Insertions can
generally not exceed 10 kbp, usually not exceed 5 kbp, generally
being at least 50 bp, more usually at least 200 bp.
[0130] The region(s) of homology can include mutations, where
mutations can further inactivate the target gene, in providing for
a frame shift, or changing a key amino acid, or the mutation can
correct a dysfunctional allele, etc. Usually, the mutation can be a
subtle change, not exceeding about 5% of the homologous flanking
sequences.
[0131] The construct can be prepared in accordance with methods
known in the art, various fragments can be brought together,
introduced into appropriate vectors, cloned, analyzed and then
manipulated further until the desired construct has been achieved
(see, for example FIGS. 5-11). Various modifications can be made to
the sequence, to allow for restriction analysis, excision,
identification of probes, etc. Silent mutations can be introduced,
as desired. At various stages, restriction analysis, sequencing,
amplification with the polymerase chain reaction, primer repair, in
vitro mutagenesis, etc. can be employed.
[0132] The construct can be prepared using a bacterial vector,
including a prokaryotic replication system, e.g. an origin
recognizable by E. coli, at each stage the construct can be cloned
and analyzed. A marker, the same as or different from the marker to
be used for insertion, can be employed, which can be removed prior
to introduction into the target cell. Once the vector containing
the construct has been completed, it can be further manipulated,
such as by deletion of the bacterial sequences, linearization,
introducing a short deletion in the homologous sequence. After
final manipulation, the construct can be introduced into the
cell.
[0133] Techniques which can be used to allow the DNA construct
entry into the host cell include calcium phosphate/DNA
co-precipitation, microinjection of DNA into the nucleus,
electroporation, bacterial protoplast fusion with intact cells,
transfection, or any other technique known by one skilled in the
art. The DNA can be single or double stranded, linear or circular,
relaxed or supercoiled DNA. For various techniques for transfecting
mammalian cells, see, for example, Keown et al., Methods in
Enzymology Vol. 185, pp. 527-537 (1990).
[0134] The present invention further includes recombinant
constructs comprising one or more of the sequences as broadly
described above (for example in Tables 9-12). The constructs
comprise a vector, such as a plasmid or viral vector, into which a
sequence of the invention has been inserted, in a forward or
reverse orientation. The construct can also include regulatory
sequences, including, for example, a promoter, operably linked to
the sequence. Large numbers of suitable vectors and promoters are
known to those of skill in the art, and are commercially available.
The following vectors are provided by way of example: pBs, pQE-9
(Qiagen), phagescript, PsiX174, pBluescript SK, pBsKS, pNH8a,
pNH16a, pNH18a, pNH46a (Stratagene); pTrc99A, pKK223-3, pKK233-3,
pDR540, pRIT5 (Pharmacia). Eukaryotic: pWLneo, pSv2cat, pOG44,
pXT1, pSG (Stratagene) pSVK3, pBPv, pMSG, pSVL (Pharmiacia). Also,
any other plasmids and vectors can be used as long as they are
replicable and viable in the host. Vectors known in the art and
those commercially available (and variants or derivatives thereof)
can in accordance with the invention be engineered to include one
or more recombination sites for use in the methods of the
invention. Such vectors can be obtained from, for example, Vector
Laboratories Inc., Invitrogen, Promega, Novagen, NEB, Clontech,
Boehringer Mannheim, Pharmacia, EpiCenter, OriGenes Technologies
Inc., Stratagene, PerkinElmer, Pharmingen, and Research Genetics.
Other vectors of interest include eukaryotic expression vectors
such as pFastBac, pFastBacHT, pFastBacDUAL, pSFV, and pTet-Splice
(Invitrogen), pEUK-C1, pPUR, pMAM, pMAMneo, pB1101, pB1121, pDR2,
pCMVEBNA, and pYACneo (Clontech), pSVK3, pSVL, pMSG, pCH110, and
pKK232-8 (Pharmacia, Inc.), p3'SS, pXT1, pSG5, pPbac, pMbac,
pMClneo, and pOG44 (Stratagene, Inc.), and pYES2, pAC360,
pBlueBacHis A, B, and C, pVL1392, pBlueBacIII, pCDM8, pcDNA1,
pZeoSV, pcDNA3 pREP4, pCEP4, and pEBVHis (Invitrogen, Corp.) and
variants or derivatives thereof.
[0135] Other vectors suitable for use in the invention include
pUC18, pUC19, pBlueScript, pSPORT, cosmids, phagemids, YAC's (yeast
artificial chromosomes), BAC's (bacterial artificial chromosomes),
P1 (Escherichia coli phage), pQE70, pQE60, pQE9 (quagan), pBS
vectors, PhageScript vectors, BlueScript vectors, pNH8A, pNH16A,
pNH18A, pNH46A (Stratagene), pcDNA3 (Invitrogen), pGEX, pTrsfus,
pTrc99A, pET-5, pET-9, pKK223-3, pKK233-3, pDR540, pRIT5
(Pharmacia), pSPORT1, pSPORT2, pCMVSPORT2.0 and pSV-SPORT1
(Invitrogen) and variants or derivatives thereof. Viral vectors can
also be used, such as lentiviral vectors (see, for example, WO
03/059923; Tiscomia et al. PNAS 100:1844-1848 (2003)).
[0136] Additional vectors of interest include pTrxFus, pThioHis,
pLEX, pTrcHis, pTrcHis2, pRSET, pBlueBacHis2, pcDNA3.1/His,
pcDNA3.1(-)/Myc-His, pSecTag, pEBVHis, pPIC9K, pPIC3.5K, pAO815,
pPICZ, pPICZA, pPICZB, pPICZC, pGAPZA, pGAPZB, pGAPZC, pBlueBac4.5,
pBlueBacHis2, pMelBac, pSinRep5, pSinHis, pIND, pIND(SP1), pVgRXR,
pcDNA2.1, pYES2, pZErO11, pZErO-2.1, pCR-Blunt, pSE280, pSE380,
pSE420, pVL1392, pVL1393, pCDM8, pcDNA1.1, pcDNA 1.1/Amp, pcDNA3.1,
pcDNA3.1/Zeo, pSe, SV2, pRc/CMV2, pRc/RSV, pREP4, pREP7, pREP8,
pREP9, pREP 10, pCEP4, pEBVHis, pCR3.1, pCR2.1, pCR3.1-Uni, and
pCRBac from Invitrogen; .lambda. ExCell, .lambda. gt11, pTrc99A,
pKK223-3, pGEX-1 .lambda. T, pGEX-2T, pGEX-2TK, pGEX-4T-1,
pGEX-4T-2, pGEX-4T-3, pGEX-3X, pGEX-5X-1, pGEX-5X-2, pGEX-5X-3,
pEZZ18, pRIT2T, pMC1871, pSVK3, pSVL, pMSG, pCH110, pKK232-8,
pSL1180, pNEO, and pUC4K from Pharmacia; pSCREEN-1b(+), pT7Blue(R),
pT7Blue-2, pCITE-4abc(+), pOCUS-2, pTAg, pET-32LIC, pET-30LIC,
pBAC-2 cp LIC, pBACgus-2 cp LIC, pT7Blue-2 LIC, pT7Blue-2, .lambda.
SCREEN-1, .lambda. BlueSTAR, pET-3abcd, pET-7abc, pET9abcd,
pET11abcd, pET12abc, pET-14b, pET-15b, pET-16b, pET-17b-pET-17xb,
pET-19b, pET-20b(+), pET-21abcd(+), pET-22b(+), pET-23abcd(+),
pET-24abcd(+), pET-25b(+), pET-26b(+), pET-27b(+), pET-28abc(+),
pET-29abc(+), pET-30abc(+), pET-31b(+), pET-32abc(+), pET-33b(+),
pBAC-1, pBACgus-1, pBAC4x-1, pBACgus4x-1, pBAC-3 cp, pBACgus-2 cp,
pBACsurf-1, pig, Signal pig, pYX, Selecta Vecta-Neo, Selecta
Vecta-Hyg, and Selecta Vecta-Gpt from Novagen; pLexA, pB42AD,
pGBT9, pAS2-1, pGAD424, pACT2, pGAD GL, pGAD GH, pGAD10, pGilda,
pEZM3, pEGFP, pEGFP-1, pEGFP--N, pEGFP--C, pEBFP, pGFPuv, pGFP,
p6xHis-GFP, pSEAP2-Basic, pSEAP2-Contral, pSEAP2-Promoter,
pSEAP2-Enhancer, p1 p.beta.gal-Basic, p.beta.gal-Control,
p.beta.gal-Promoter, p.beta.gal-Enhancer, pCMV, pTet-Off, pTet-On,
pTK-Hyg, pRetro-Off, pRetro-On, pIRES1neo, pIRES1 hyg, pLXSN,
pLNCX, pLAPSN, pMAMneo, pMAMneo-CAT, pMAMneo-LUC, pPUR, pSV2neo,
pYEX4T-1/2/3, pYEX-S1, pBacPAK-His, pBacPAK8/9, pAcUW31, BacPAK6,
pTriplEx, .lambda.gt10, .lambda.gt11, pWE15, and .lambda.TriplEx
from Clontech; Lambda ZAP II, pBK-CMV, pBK-RSV, pBluescript II KS
+/-, pBluescript 11 SK +/-, pAD-GAL4, pBD-GAL4 Cam, pSurfscript,
Lambda FIX II, Lambda DASH, Lambda EMBL3, Lambda EMBL4, SuperCos,
pCR-Scrigt Amp, pCR-Script Cam, pCR-Script Direct, pBS +/-, pBC KS
+/-, pBC SK +/-, Phagescript, pCAL-n-EK, pCAL-n, pCAL-c, pCAL-kc,
pET-3abcd, pET-1 labcd, pSPUTK, pESP-1, pCMVLacI, pOPRSVI/MCS,
pOPI3 CAT,pXT1, pSG5, pPbac, pMbac, pMClneo, pMClneo Poly A, pOG44,
pOG45, pFRT.beta.GAL, pNEOPGAL, pRS403, pRS404, pRS405, pRS406,
pRS413, pRS414, pRS415, and pRS416 from Stratagene.
[0137] Additional vectors include, for example, pPC86, pDBLeu,
pDBTrp, pPC97, p2.5, pGAD1-3, pGAD10, pACt, pACT2, pGADGL, pGADGH,
pAS2-1, pGAD424, pGBT8, pGBT9, pGAD-GAL4, pLexA, pBD-GAL4, pHISi,
pHISi-1, placZi, pB42AD, pDG202, pJK202, pJG4-5, pNLexA, pYESTrp
and variants or derivatives thereof.
[0138] Also, any other plasmids and vectors known in the art can be
used as long as they are replicable and viable in the host.
Selection of Homologously Recombined Cells
[0139] Cells that have been homologously recombined to knock-out
expression of the porcine CMP-Neu5Ac hydroxylase gene can then be
grown in appropriately-selected medium to identify cells providing
the appropriate integration. Those cells which show the desired
phenotype can then be further analyzed by restriction analysis,
electrophoresis, Southern analysis, polymerase chain reaction, or
another technique known in the art. By identifying fragments which
show the appropriate insertion at the target gene site, cells can
be identified in which homologous recombination has occurred to
inactivate or otherwise modify the target gene.
[0140] The presence of the selectable marker gene inserted into the
CMP-Neu5Ac hydroxylase gene establishes the integration of the
target construct into the host genome. Those cells which show the
desired phenotype can then be further analyzed by restriction
analysis, electrophoresis, Southern analysis, polymerase chain
reaction, monoclonal antibody assays, Fluorescent Activated Cell
Sorter (FACS), or any other techniques or methods known in the art
to analyze the DNA in order to establish whether homologous or
non-homologous recombination occurred. This can be determined by
employing probes for the insert and then sequencing the 5' and 3'
regions flanking the insert for the presence of the CMP-Neu5Ac
hydroxylase gene extending beyond the flanking regions of the
construct or identifying the presence of a deletion, when such
deletion is introduced. Primers can also be used which are
complementary to a sequence within the construct and complementary
to a sequence outside the construct and at the target locus. In
this way, one can only obtain DNA duplexes having both of the
primers present in the complementary chains if homologous
recombination has occurred. By demonstrating the presence of the
primer sequences or the expected size sequence, the occurrence of
homologous recombination is supported.
[0141] The polymerase chain reaction used for screening homologous
recombination events is described, for example, in Kim and
Smithies, Nucleic Acids Res. 16:8887-8903, 1988; and Joyner, et
al., Nature 338:153-156, 1989.
[0142] An alternative method for screening homologous recombination
events includes utilizing monoclonal or polyclonal antibodies
specific for porcine CMP-Neu5Ac Hydroxylase and/or NeuSGc, as
described in, for example, Malykh, et al., European Journal of Cell
Biology 80, 48-58 (2001), Malykh, et al., Glycoconjugate J. 15,
885-893 (1998).
[0143] Further characterization of porcine cells lacking expression
of functional CMP-Neu5Ac Hydroxylase due to homologous
recombination events include, but are not limited to, Southern Blot
analysis, Northern Blot analysis, specific lectin binding assays,
and/or sequence analysis, or by using anti-Neu5Gc or
anti-CMP-Neu5Ac hydroxylase antibody assays as described, for
example, in Y. Malykh, et. al. Biochem J. 370: 601-607 (2003); Y.
Malykh, et al. European Journal of Cell Biology 80: 48-58 (2001);
Y. Malykh et al. Glycoconjugate J. 15: 885-893 (1998). See
generally, for example, A. Sharma, et al. Transplantation 75(4):
430-436 (2003).
[0144] The cell lines obtained from the first round of targeting
are likely to be heterozygous for the targeted allele.
Homozygosity, in which both alleles are modified, can be achieved
in a number of ways. One approach is to grow up a number of cells
in which one copy has been modified and then to subject these cells
to another round of targeting of the remaining porcine CMP-Neu5Ac
hydroxylase allele using a different selectable marker.
Alternatively, homozygotes can be obtained by breeding animals
heterozygous for the modified allele, according to traditional
Mendelian genetics. In some situations, it can be desirable to have
two different modified alleles. This can be achieved by successive
rounds of gene targeting or by breeding heterozygotes, each of
which carries one of the desired modified alleles.
VIII. Genetic Manipulation of Additional Genes to Overcome
Immunologic Barriers of Xenotransplantation
[0145] In one aspect of the invention, cells homozygous for the
nonfunctional CMP-Neu5Ac hydroxylase gene can be subject to further
genetic modification. For example, one can introduce additional
genetic capability into the homozygotic hosts, where the endogenous
CMP Neu5Ac hydroxylase alleles have been made nonfunctional, to
substitute, replace or provide different genetic capability to the
host. One can remove the marker gene after homogenotization. By
introducing a construct comprising substantially the same
homologous DNA, possibly with extended sequences, having the marker
gene portion of the original construct deleted, one can be able to
obtain homologous recombination with the target locus. By using a
combination of marker genes for integration, one providing positive
selection and the other negative selection, in the removal step,
one can select against the cells retaining the marker genes.
[0146] In one embodiment, porcine cells are provided that lack the
CMP-Neu5Ac hydroxylase gene and the .alpha.(1,3)GT gene. Animals
lacking functional CMP-Neu5Ac hydroxylase can be produced according
to the present invention, and then cells from this animal can be
used to knockout the .alpha.(1,3)GT gene. Homozygous .alpha.(1,3)GT
negative porcine have recently been reported (Phelps et. al.
Science 2003; WO 04/028243). Alternatively, cells from these
.alpha.(1,3)GT knockout animals can be used and further modified to
inactivate the CMP-Neu5Ac hydroxylase gene.
[0147] In another embodiment, porcine cells are also provided that
lack the porcine CMP-Neu5Ac hydroxylase gene and produce human
complement inhibiting proteins. Animals lacking functional porcine
CMP-Neu5Ac hydroxylase gene can be produced according to the
present invention, and then cells from this animal can be further
modified to express human complement inhibiting proteins, such as,
but not limited to, CD59 (cDNA reported by Philbrick, W. M., et al.
(1990) Eur. J. Immunol. 20:87-92), human decay accelerating factor
(DAF)(cDNA reported by Medof, et al. (1987) Proc. Natl. Acad. Sci.
USA 84: 2007), and human membrane cofactor protein (MCP) (cDNA
reported by Lublin, D., et al. (1988) J. Exp. Med. 168:
181-194).
[0148] In an alternative embodiment, cells from transgenic pigs
producing human complement inhibiting proteins can be used and
further modified to inactivate the porcine CMP-Neu5Ac hydroxylase
gene. Transgenic pigs producing human complement inhibiting
proteins are known in the art (see, for example, U.S. Pat. No.
6,166,288).
[0149] In a further embodiment, porcine cells are provided that
lack the porcine CMP-Neu5Ac hydroxylase gene and the porcine
Forssman synthetase (FSM) gene. Animals lacking functional porcine
CMP-Neu5Ac hydroxylase gene can be produced according to the
present invention, and then cells from this animal can be further
modified to knockout the porcine FSM synthetase gene, which is
involved in the production of gal-.alpha.-gal epitopes, and plays a
role in xenotransplant rejection. The porcine FSM synthetase gene
has recently been identified (see U.S. Application 60/568,922).
Alternatively, cells from these FSM synthetase gene knockout
animals can be used and further modified to inactivate the porcine
CMP-Neu5Ac hydroxylase gene.
[0150] In a still further embodiment, porcine cells are provided
that lack the porcine CMP-Neu5Ac hydroxylase gene and the porcine
isogloboside 3 synthase gene. Animals lacking functional porcine
CMP-Neu5Ac hydroxylase gene can be produced according to the
present invention, and then cells from this animal can be used to
knockout the porcine iGb3 synthase gene. The porcine iGb3 synthase
gene has recently been reported (U.S. Application No. 60/517,524).
Alternatively, cells from these porcine iGb3 synthase gene knockout
animals can be used and further modified to inactivate the porcine
CMP-Neu5Ac hydroxylase gene.
[0151] In another embodiment, porcine cells are provided that lack
the porcine CMP-Neu5Ac hydroxylase gene, the .alpha.(1,3)GT gene,
the FSM synthetase gene, and the porcine iGb3 synthase gene.
Animals lacking functional CMP-Neu5Ac hydroxylase gene can be
produced according to the present invention, and then cells from
this animal can be used to knockout the .alpha.(1,3)GT gene, the
FSM synthetase gene, and the porcine iGb3 synthase gene. Homozygous
.alpha.(1,3)GT-negative porcine have recently been reported (Phelps
et al. supra, Science 2003; WO 04/028243) Alternatively, cells from
these .alpha.(1,3)GT knockout animals can be used and further
modified to inactivate the porcine iGb3 synthase gene, the porcine
FSM synthetase gene, and the CMP-Neu5Ac hydroxylase gene, and, in
addition, express human complement inhibiting proteins, such as,
but not limited to, CD59, human decay accelerating factor (DAF),
and human membrane cofactor protein (MCP).
VIII. Production of Genetically Modified Animals
[0152] The present invention provides methods of producing a
transgenic pig that lacks expression of CMP-Neu5Ac hydroxylase
through the genetic modification of porcine totipotent embryonic
cells. In one embodiment, the animals can be produced by: (a)
identifying one or more target CMP-Neu5Ac hydroxylase nucleic acid
genomic sequences in an animal; (b) preparing one or more
homologous recombination vectors targeting the CMP-Neu5Ac
hydroxylase nucleic acid genomic sequences; (c) inserting the one
or more targeting vectors into the genomes of a plurality of
totipotent cells of the animal, thereby producing a plurality of
transgenic totipotent cells; (d) obtaining a tetraploid blastocyst
of the animal; (e) inserting the plurality of totipotent cells into
the tetraploid blastocyst, thereby producing a transgenic embryo;
(f) transferring the embryo to a recipient female animal; and (g)
allowing the embryo to develop to term in the female animal. The
method of transgenic animal production described here by which to
generate a transgenic pig is further generally described in U.S.
Pat. No. 6,492,575.
[0153] In another embodiment, the totipotent cells can be embryonic
stem (ES) cells. The isolation of ES cells from blastocysts, the
establishing of ES cell lines and their subsequent cultivation are
carried out by conventional methods as described, for example, by
Doetchmann et al., J. Embryol. Exp. Morph. 87:27-45 (1985); Li et
al., Cell 69:915-926 (1992); Robertson, E. J. "Tetracarcinomas and
Embryonic Stem Cells: A Practical Approach," ed. E. J. Robertson,
IRL Press, Oxford, England (1987); Wurst and Joyner, "Gene
Targeting: A Practical Approach," ed. A. L. Joyner, IRL Press,
Oxford, England (1993); Hogen et al., "Manipulating the Mouse
Embryo: A Laboratory Manual," eds. Hogan, Beddington, Costantini
and Lacy, Cold Spring Harbor Laboratory Press, New York (1994); and
Wang, et al., Nature 336:741-744 (1992). For example, after
transforming embryonic stem cells with the targeting vector to
alter the CMP-Neu5Ac hydroxylase gene, the cells can be plated onto
a feeder layer in an appropriate medium, for example, such as fetal
bovine serum enhanced DMEM. Cells containing the construct can be
detected by employing a selective medium, and after sufficient time
for colonies to grow, colonies can be picked and analyzed for the
occurrence of homologous recombination. Polymerase chain reaction
can be used, with primers within and without the construct sequence
but at the target locus. Those colonies which show homologous
recombination can then be used for embryo manipulating and
blastocyst injection. Blastocysts can be obtained from
superovulated females. The embryonic stem cells can then be
trypsinized and the modified cells added to a droplet containing
the blastocysts. At least one of the modified embryonic stem cells
can be injected into the blastocoel of the blastocyst. After
injection, at least one of the blastocysts can be returned to each
uterine horn of pseudopregnant females. Females are then allowed to
go to term and the resulting litters screened for mutant cells
having the construct. The blastocysts are selected for different
parentage from the transformed ES cells. By providing for a
different phenotype of the blastocyst and the ES cells, chimeric
progeny can be readily detected, and then genotyping can be
conducted to probe for the presence of the modified CMP-Neu5Ac
hydroxylase gene.
[0154] In a further embodiment of the invention, the totipotent
cells can be embryonic germ (EG) cells. Embryonic Germ cells are
undifferentiated cells functionally equivalent to ES cells, that is
they can be cultured and transfected in vitro, then contribute to
somatic and germ cell lineages of a chimera (Stewart et al., Dev.
Biol. 161:626-628 (1994)). EG cells are derived by culture of
primordial germ cells, the progenitors of the gametes, with a
combination of growth factors: leukemia inhibitory factor, steel
factor and basic fibroblast growth factor (Matsui, et al., Cell
70:841-847 (1992); Resnick, et al., Nature 359:550-551 (1992)). The
cultivation of EG cells can be carried out using methods known to
one skilled in the art, such as described in Donovan et al.,
"Transgenic Animals, Generation and Use," Ed. L. M. Houdebine,
Harwood Academic Publishers (1997).
[0155] Tetraploid blastocysts for use in the invention can be
obtained by natural zygote production and development, or by known
methods by electrofusion of two-cell embryos and subsequently
cultured as described, for example, by James, et al., Genet. Res.
Camb. 60:185-194 (1992); Nagy and Rossant, "Gene Targeting: A
Practical Approach," ed. A. L. Joyner, IRL Press, Oxford, England
(1993); or by Kubiak and Tarkowski, Exp. Cell Res. 157:561-566
(1985).
[0156] The introduction of the ES cells or EG cells into the
blastocysts can be carried out by any method known in the art, for
example, as described by Wang, et al., EMBO J. 10:2437-2450
(1991).
[0157] A "plurality" of totipotent cells can encompass any number
of cells greater than one. For example, the number of totipotent
cells for use in the present invention can be about 2 to about 30
cells, about 5 to about 20 cells, or about 5 to about 10 cells. In
one embodiment, about 5-10 ES cells taken from a single cell
suspension are injected into a blastocyst immobilized by a holding
pipette in a micromanipulation apparatus. Then the embryos are
incubated for at least 3 hours, possibly overnight, prior to
introduction into a female recipient animal via methods known in
the art (see for example Robertson, E. J. "Teratocarcinomas and
Embryonic Stem Cells: A Practical Approach" IRL Press, Oxford,
England (1987)). The embryo can then be allowed to develop to term
in the female animal.
Somatic Cell Nuclear Transfer to Produce Cloned, Transgenic
Offspring
[0158] The present invention provides a method for cloning a pig
lacking a functional CMP-Neu5Ac hydroxylase gene via somatic cell
nuclear transfer. In general, a wide variety of methods to
accomplish mammalian cloning are currently being rapily developed
and reported, any method that accomplishes the desired result can
be used in the present invention. Nonlimiting examples of such
methods are described below. For example, the pig can be produced
by a nuclear transfer process comprising the following steps:
obtaining desired differentiated pig cells to be used as a source
of donor nuclei; obtaining oocytes from a pig; enucleating the
oocytes; transferring the desired differentiated cell or cell
nucleus into the enucleated oocyte, e.g., by fusion or injection,
to form NT units; activating the resultant NT unit; and
transferring said cultured NT unit to a host pig such that the NT
unit develops into a fetus.
[0159] Nuclear transfer techniques or nuclear transplantation
techniques are known in the art (Campbell et al, Theriogenology,
43:181 (1995); Collas, et al, Mol. Report Dev., 38:264-267 (1994);
Keefer et al, Biol. Reprod., 50:935-939 (1994); Sims, et al, Proc.
Natl. Acad. Sci., USA, 90:6143-6147 (1993); WO 94/26884; WO
94/24274, and WO 90/03432, U.S. Pat. Nos. 4,944,384 and 5,057,420).
In one nonlimiting example, methods are provided such as those
described in U.S. Patent Publication No. 2003/0046722 to Collas, et
al., which describes methods for cloning mammals that allow the
donor chromosomes or donor cells to be reprogrammed prior to
insertion into an enucleated oocyte. The invention also describes
methods of inserting or fusing chromosomes, nuclei or cells with
oocytes.
[0160] A donor cell nucleus, which has been modified to alter the
CMP-Neu5Ac hydroxylase gene, is transferred to a recipient porcine
oocyte. The use of this method is not restricted to a particular
donor cell type. The donor cell can be as described in Wilmut, et
al., Nature 385 810 (1997); Campbell, et al., Nature 380 64-66
(1996); or Cibelli, et al., Science 280 1256-1258 (1998). All cells
of normal karyotype, including embryonic, fetal and adult somatic
cells which can be used successfully in nuclear transfer can in
principle be employed. Fetal fibroblasts are a particularly useful
class of donor cells. Generally suitable methods of nuclear
transfer are described in Campbell, et al., Theriogenology 43 181
(1995), Collas, et al., Mol. Reprod Dev. 38 264-267 (1994), Keefer,
et al., Biol. Reprod. 50 935-939 (1994), Sims, et al., Proc. Nat'l.
Acad. Sci. USA 90 6143-6147 (1993), WO-A-9426884, WO-A-9424274,
WO-A-9807841, WO-A-9003432, U.S. Pat. No. 4,994,384 and U.S. Pat.
No. 5,057,420. Differentiated or at least partially differentiated
donor cells can also be used. Donor cells can also be, but do not
have to be, in culture and can be quiescent. Nuclear donor cells
which are quiescent are cells which can be induced to enter
quiescence or exist in a quiescent state in vivo. Prior art methods
have also used embryonic cell types in cloning procedures
(Campbell, et al. (Nature, 380:64-68, 1996) and Stice, et al (Biol.
Reprod., 20 54:100-110, 1996).
[0161] Somatic nuclear donor cells may be obtained from a variety
of different organs and tissues such as, but not limited to, skin,
mesenchyme, lung, pancreas, heart, intestine, stomach, bladder,
blood vessels, kidney, urethra, reproductive organs, and a
disaggregated preparation of a whole or part of an embryo, fetus or
adult animal. In a suitable embodiment of the invention, nuclear
donor cells are selected from the group consisting of epithelial
cells, fibroblast cells, neural cells, keratinocytes, hematopoietic
cells, melanocytes, chondrocytes, lymphocytes (B and T),
macrophages, monocytes, mononuclear cells, cadiac muscle cells,
other muscle cells, granulose cells, cumulus cells, epidermal cells
or endothelial cells. In another embodiment, the nuclear cell is an
embryonic stem cell. In a preferred embodiment, fibroblast cells
can be used as donor cells.
[0162] In another embodiment of the invention, the nuclear donor
cells of the invention are germ cells of an animal. Any germ cell
of an animal species in the embryonic, fetal, or adult stage may be
used as a nuclear donor cell. In a suitable embodiment, the nuclear
donor cell is an embryonic germ cell.
[0163] Nuclear donor cells may be arrested in any phase of the cell
cycle (GO, GI, G2, S, M) so as to ensure coordination with the
acceptor cell. Any method known in the art may be used to
manipulate the cell cycle phase. Methods to control the cell cycle
phase include, but are not limited to, GO quiescence induced by
contact inhibition of cultured cells, GO quiescence induced by
removal of serum or other essential nutrient, GO quiescence induced
by senescence, GO quiescence induced by addition of a specific
growth factor; GO or GI quiescence induced by physical or chemical
means such as heat shock, hyperbaric pressure or other treatment
with a chemical, hormone, growth factor or other substance; S-phase
control via treatment with a chemical agent which interferes with
any. Point of the replication procedure; M-phase control via
selection using fluorescence activated cell sorting, mitotic shake
off, treatment with microtubule disrupting agents or any chemical
which disrupts progression in mitosis (see also Freshney, R. I,.
"Culture of Animal Cells: A Manual of Basic Technique," Alan R.
Liss, Inc, New York (1983).
[0164] Methods for isolation of oocytes are well known in the art.
Essentially, this can comprise isolating oocytes from the ovaries
or reproductive tract of a pig. A readily available source of pig
oocytes is slaughterhouse materials. For the combination of
techniques such as genetic engineering, nuclear transfer and
cloning, oocytes must generally be matured in vitro before these
cells can be used as recipient cells for nuclear transfer, and
before they can be fertilized by the sperm cell to develop into an
embryo. This process generally requires collecting immature
(prophase I) oocytes from mammalian ovaries, e.g., bovine ovaries
obtained at a slaughterhouse, and maturing the oocytes in a
maturation medium prior to fertilization or enucleation until the
oocyte attains the metaphase II stage, which in the case of bovine
oocytes generally occurs about 18-24 hours post-aspiration. This
period of time is known as the "maturation period".
[0165] A metaphase II stage oocyte can be the recipient oocyte, at
this stage it is believed that the oocyte can be or is sufficiently
"activated" to treat the introduced nucleus as it does a
fertilizing sperm. Metaphase II stage oocytes, which have been
matured in vivo have been successfully used in nuclear transfer
techniques. Essentially, mature metaphase II oocytes can be
collected surgically from either non-superovulated or superovulated
porcine 35 to 48, or 39-41, hours past the onset of estrus or past
the injection of human chorionic gonadotropin (hCG) or similar
hormone.
[0166] After a fixed time maturation period, which ranges from
about 10 to 40 hours, and preferably about 16-18 hours, the oocytes
can be enucleated. Prior to enucleation the oocytes can be removed
and placed in appropriate medium, such as HECM containing 1
milligram per milliliter of hyaluronidase prior to removal of
cumulus cells. The stripped oocytes can then be screened for polar
bodies, and the selected metaphase II oocytes, as determined by the
presence of polar bodies, are then used for nuclear transfer.
Enucleation follows.
[0167] Enucleation can be performed by known methods, such as
described in U.S. Pat. No. 4,994,384. For example, metaphase II
oocytes can be placed in either HECM, optionally containing 7.5
micrograms per milliliter cytochalasin B, for immediate
enucleation, or can be placed in a suitable medium, for example an
embryo culture medium such as CR1 aa, plus 10% estrus cow serum,
and then enucleated later, preferably not more than 24 hours later,
and more preferably 16-18 hours later.
[0168] Enucleation can be accomplished microsurgically using a
micropipette to remove the polar body and the adjacent cytoplasm.
The oocytes can then be screened to identify those of which have
been successfully enucleated. One way to screen the oocytes is to
stain the oocytes with 1 microgram per milliliter 33342 Hoechst dye
in HECM, and then view the oocytes under ultraviolet irradiation
for less than 10 seconds. The oocytes that have been successfully
enucleated can then be placed in a suitable culture medium, for
example, CR1 aa plus 10% serum.
[0169] A single mammalian cell of the same species as the
enucleated oocyte can then be transferred into the perivitelline
space of the enucleated oocyte used to produce the NT unit. The
mammalian cell and the enucleated oocyte can be used to produce NT
units according to methods known in the art. For example, the cells
can be fused by electrofusion. Electrofusion is accomplished by
providing a pulse of electricity that is sufficient to cause a
transient breakdown of the plasma membrane. This breakdown of the
plasma membrane is very short because the membrane reforms rapidly.
Thus, if two adjacent membranes are induced to breakdown and upon
reformation the lipid bilayers intermingle, small channels can open
between the two cells. Due to the thermodynamic instability of such
a small opening, it enlarges until the two cells become one. See,
for example, U.S. Pat. No. 4,997,384 by Prather et al. A variety of
electrofusion media can be used including, for example, sucrose,
mannitol, sorbitol and phosphate buffered solution. Fusion can also
be accomplished using Sendai virus as a fusogenic agent (Graham,
Wister Inot. Symp. Monogr., 9, 19, 1969). Also, the nucleus can be
injected directly into the oocyte rather than using electroporation
fusion. See, for example, Collas and Barnes, Mol. Reprod. Dev.,
38:264-267 (1994). After fusion, the resultant fused NT units are
then placed in a suitable medium until activation, for example,
CR1aa medium. Typically activation can be effected shortly
thereafter, for example less than 24 hours later, or about 4-9
hours later.
[0170] The NT unit can be activated by any method that accomplishes
the desired result. Such methods include, for example, culturing
the NT unit at sub-physiological temperature, in essence by
applying a cold, or actually cool temperature shock to the NT unit.
This can be most conveniently done by culturing the NT unit at room
temperature, which is cold relative to the physiological
temperature conditions to which embryos are normally exposed.
Alternatively, activation can be achieved by application of known
activation agents. For example, penetration of oocytes by sperm
during fertilization has been shown to activate prefusion oocytes
to yield greater numbers of viable pregnancies and multiple
genetically identical pigs after nuclear transfer. Also, treatments
such as electrical and chemical shock can be used to activate NT
embryos after fusion. See, for example, U.S. Pat. No. 5,496,720, to
Susko-Parrish, et al. Additionally, activation can be effected by
simultaneously or sequentially by increasing levels of divalent
cations in the oocyte, and reducing phosphorylation of cellular
proteins in the oocyte. This can generally be effected by
introducing divalent cations into the oocyte cytoplasm, e.g.,
magnesium, strontium, barium or calcium, e.g., in the form of an
ionophore. Other methods of increasing divalent cation levels
include the use of electric shock, treatment with ethanol and
treatment with caged chelators. Phosphorylation can be reduced by
known methods, for example, by the addition of kinase inhibitors,
e.g., serine-threonine kinase inhibitors, such as
6-dimethyl-aminopurine, staurosporine, 2-aminopurine, and
sphingosine. Alternatively, phosphorylation of cellular proteins
can be inhibited by introduction of a phosphatase into the oocyte,
e.g., phosphatase 2A and phosphatase 2B.
[0171] The activated NT units can then be cultured in a suitable in
vitro culture medium until the generation of cell colonies. Culture
media suitable for culturing and maturation of embryos are well
known in the art. Examples of known media, which can be used for
embryo culture and maintenance, include Ham's F-10+10% fetal calf
serum (FCS), Tissue Culture Medium-199 (TCM-199)+10% fetal calf
serum, Tyrodes-Albumin-Lactate-Pyruvate (TALP), Dulbecco's
Phosphate Buffered Saline (PBS), Eagle's and Whitten's media.
[0172] Afterward, the cultured NT unit or units can be washed and
then placed in a suitable media contained in well plates which
preferably contain a suitable confluent feeder layer. Suitable
feeder layers include, by way of example, fibroblasts and
epithelial cells. The NT units are cultured on the feeder layer
until the NT units reach a size suitable for transferring to a
recipient female, or for obtaining cells which can be used to
produce cell colonies. Preferably, these NT units can be cultured
until at least about 2 to 400 cells, more preferably about 4 to 128
cells, and most preferably at least about 50 cells.
[0173] Activated NT units can then be transferred (embryo
transfers) to the oviduct of an female pigs. In one embodiment, the
female pigs can be an estrus-synchronized recipient gilt. Crossbred
gilts (large white/Duroc/Landrace) (280-400 lbs) can be used. The
gilts can be synchronized as recipient animals by oral
administration of 18-20 mg ReguMate (Altrenogest, Hoechst, Warren,
N.J.) mixed into the feed. Regu-Mate can be fed for 14 consecutive
days. One thousand units of Human Chorionic Gonadotropin (hCG,
Intervet America, Millsboro, Del.) can then be administered i.m.
about 105 h after the last Regu-Mate treatment. Embryo transfers of
the can then be performed about 22-26 h after the hCG injection. In
one embodiment, the pregnancy can be brought to term and result in
the birth of live offspring. In another embodiment, the pregnancy
can be 5 terminated early and embryonic cells can be harvested.
[0174] The methods for embryo transfer and recipient animal
management in the present invention are standard procedures used in
the embryo transfer industry. Synchronous transfers are important
for success of the present invention, i.e., the stage of the NT
embryo is in synchrony with the estrus cycle of the recipient
female. See, for example, Siedel, G. E., Jr. "Critical review of
embryo transfer procedures with cattle" in Fertilization and
Embryonic Development in Vitro (1981) L. Mastroianni, Jr. and J. D.
Biggers, ed., Plenum Press, New York, N.Y., page 323.
VIII. Porcine Animals, Organs, Tissues, Cells and Cell Lines
[0175] The present invention provides viable porcine in which both
alleles of the CMP-Neu5Ac hydroxylase gene have been inactivated.
The invention also provides organs, tissues, and cells derived from
such porcine, which are useful for xenotransplantation.
[0176] In one embodiment, the invention provides porcine organs,
tissues and/or purified or substantially pure cells or cell lines
obtained from pigs that lack any expression of functional
CMP-Neu5Ac hydroxylase.
[0177] In one embodiment, the invention provides organs that are
useful for xenotransplantation. Any porcine organ can be used,
including, but not limited to: brain, heart, lungs, glands, brain,
eye, stomach, spleen, pancreas, kidneys, liver, intestines, uterus,
bladder, skin, hair, nails, ears, nose, mouth, lips, gums, teeth,
tongue, salivary glands, tonsils, pharynx, esophagus, large
intestine, small intestine, rectum, anus, pylorus, thyroid gland,
thymus gland, suprarenal capsule, bones, cartilage, tendons,
ligaments, skeletal muscles, smooth muscles, blood vessels, blood,
spinal cord, trachea, ureters, urethra, hypothalamus, pituitary,
adrenal glands, ovaries, oviducts, uterus, vagina, mammary glands,
testes, seminal vesicles, penis, lymph, lymph nodes and lymph
vessels.
[0178] In another embodiment, the invention provides tissues that
are useful for xenotransplantation. Any porcine tissue can be used,
including, but not limited to: epithelium, connective tissue,
blood, bone, cartilage, muscle, nerve, adenoid, adipose, areolar,
bone, brown adipose, cancellous, muscle, cartaginous, cavernous,
chondroid, chromaffin, dartoic, elastic, epithelial, fatty,
fibrohyaline, fibrous, Gaingee, gelatinous, granulation,
gut-associated lymphoid, Haller's vascular, hard hemopoietic,
indifferent, interstitial, investing, islet, lymphatic, lymphoid,
mesenchymal, mesonephric, mucous connective, multilocular adipose,
myeloid, nasion soft, nephrogenic, nodal, osseous, osteogenic,
osteoid, periapical, reticular, retiform, rubber, skeletal muscle,
smooth muscle, and subcutaneous tissue.
[0179] In a further embodiment, the invention provides cells and
cell lines from porcine animals that lack expression of functional
alphal,3GT. In one embodiment, these cells or cell lines can be
used for xenotransplantation. Cells from any porcine tissue or
organ can be used, including, but not limited to: epithelial cells,
fibroblast cells, neural cells, keratinocytes, hematopoietic cells,
melanocytes, chondrocytes, lymphocytes (B and T), macrophages,
monocytes, mononuclear cells, cardiac muscle cells, other muscle
cells, .quadrature.hosphate cells, cumulus cells, epidermal cells,
endothelial cells, Islets of Langerhans cells, pancreatic insulin
secreting cells, pancreatic alpha-2 cells, pancreatic beta cells,
pancreatic alpha-1 cells, blood cells, blood precursor cells, bone
cells, bone precursor cells, neuronal stem cells, primordial stem
cells., hepatocytes, keratinocytes, umbilical vein endothelial
cells, aortic endothelial cells, microvascular endothelial cells,
fibroblasts, liver stellate cells, aortic smooth muscle cells,
cardiac myocytes, neurons, Kupffer cells, smooth muscle cells,
Schwann cells, and epithelial cells, erythrocytes, platelets,
neutrophils, lymphocytes, monocytes, eosinophils, basophils,
adipocytes, chondrocy-tes, pancreatic islet cells, thyroid cells,
parathyroid cells, parotid cells, tumor cells, glial cells,
astrocytes, red blood cells, white blood cells, macrophages,
epithelial cells, somatic cells, pituitary cells, adrenal cells,
hair cells, bladder cells, kidney cells, retinal cells, rod cells,
cone cells, heart cells, pacemaker cells, spleen cells, antigen
presenting cells, memory cells, T cells, B cells, plasma cells,
muscle cells, ovarian cells, uterine cells, prostate cells, vaginal
epithelial cells, sperm cells, testicular cells, germ cells, egg
cells, leydig cells, peritubular cells, sertoli cells, lutein
cells, cervical cells, endometrial cells, mammary cells, follicle
cells, mucous cells, ciliated cells, nonkeratinized epithelial
cells, keratinized epithelial cells, lung cells, goblet cells,
columnar epithelial cells, dopaminergic cells, squamous epithelial
cells, osteocytes, osteoblasts, osteoclasts, embryonic stem cells,
fibroblasts and fetal fibroblasts. In a specific embodiment,
pancreatic cells, including, but not limited to, Islets of
Langerhans cells, insulin secreting cells, 48 alpha-2 cells, beta
cells, alpha-i cells from pigs that lack expression of functional
alpha-1,3-GT are provided.
[0180] Nonviable derivatives include tisssues stripped of viable
cells by enzymatic or chemical treatment these tissue derivatives
can be further processed via crosslinking or other chemical
treatments prior to use in transplantation. In a preferred
embodiment, the derivatives include extracellular matrix derived
from a variety of tissues, including skin, urinary, bladder or
organ submucosal tissues. Also, tendons, joints and bones stripped
of viable tissue to include heart valves and other nonviable
tissues as medical devices are provided.
Therapeutic Uses
[0181] The cells can be administered into a host in order in a wide
variety of ways. Preferred modes of administration are parenteral,
intraperitoneal, intravenous, intradermal, epidural, intraspinal,
intrastemal, intra-articular, intra-synovial, intrathecal,
intra-arterial, intracardiac, intramuscular, intranasal,
subcutaneous, intraorbital, intracapsular, topical, transdermal
patch, via rectal, vaginal or urethral administration including via
suppository, percutaneous, nasal spray, surgical implant, internal
surgical paint, infusion pump, or via catheter. In one embodiment,
the agent and carrier are administered in a slow release
formulation such as a direct tissue injection or bolus, implant,
microparticle, microsphere, nanoparticle or nanosphere.
[0182] Disorders that can be treated by infusion of the disclosed
cells include, but are not limited to, diseases resulting from a
failure of a dysfunction of normal blood cell production and
maturation (i.e., aplastic anemia and hypoproliferative stem cell
disorders); neoplastic, malignant diseases in the hematopoietic
organs (e.g., leukemia and lymphomas); broad spectrum malignant
solid tumors of non-hematopoietic origin; autoimmune conditions;
and genetic disorders. Such disorders include, but are not limited
to diseases resulting from a failure or dysfunction of normal blood
cell production and maturation hyperproliferative stem cell
disorders, including aplastic anemia, pancytopenia,
agranulocytosis, thrombocytopenia, red cell aplasia, Blackfan
Diamond syndrome, due to drugs, radiation, or infection,
idiopathic; hematopoietic malignancies including acute
lymphoblastic (lymphocytic) leukemia, chronic lymphocytic leukemia,
acute myelogenous leukemia, chronic myelogenous, leukemia, acute
malignant myelosclerosis, multiple myeloma, polycythemia vera,
agnogenic myelometaplasia, Waldenstrom's macroglobulinemia,
Hodgkin's lymphoma, non-Hodgkin's lymphoma; immunosuppression in
patients with malignant, solid tumors including malignant melanoma,
carcinoma of the stomach, ovarian carcinoma, breast carcinoma,
small cell lung carcinoma, retinoblastoma, testicular carcinoma,
glioblastoma, rhabdomyosarcoma, neuroblastoma, Ewing's sarcoma,
lymphonia; autoinimune diseases including rheumatoid arthritis,
diabetes type 1, chronic hepatitis, multiple sclerosis, systemic
lupus erythematosus; genetic (congenital) disorders including
anemias, familial aplastic, Fanconi's syndrome, dihydrofolate
reductase deficiencies, formamino transferase deficiency,
Lesch-Nyhan syndrome, congenital dyserythropoietic syndrome IIV,
Chwachmann-Diamond syndrome, dihydrofolate reductase deficiencies,
forinamino transferase deficiency, Lesch-Nyhan syndrome, congenital
spherocytosis, congenital elliptocytosis, congenital
stomatocytosis, congenital Rh null disease, paroxysmal nocturnal
hemoglobinuria, G6PD (glucose [ ]hosphate dehydrogenase) variants
1, 2, 3, pyruvate kinase deficiency, congenital erythropoietin
sensitivity, deficiency, sickle cell disease and trait,
thalassernia alpha, beta, gamma, met-hemoglobinemia, congenital
disorders of immunity, severe combined immunodeficiency disease
(SCID), bare lymphocyte syndrome, ionophore-responsive combined
immunodeficiency, combined immunodeficiency with a capping
abnormality, nucleoside phosphorylase deficiency, granulocyte actin
deficiency, infantile agranulocytosis, Gaucher's disease, adenosine
deaminase deficiency, Kostmann's syndrome, reticular dysgenesis,
congenital Leukocyte dysfunction syndromes; and others such as
osteoporosis, myeloselerosis, acquired hemolytic anemias, acquired
immunodeficiencies, infectious disorders causing primary or
secondary immunodeficiencies, bacterial infections (e.g.,
Brucellosis, Listerosis, tuberculosis, leprosy), parasitic
infections (e.g., malaria, Leishmaniasis), fungal infections,
disorders involving disproportionsin lymphoid cell sets and
impaired immune functions due to aging, phagocyte disorders,
Kostmann's agranulocytosis, chronic granulomatous disease,
Chediak-Higachi syndrome, neutrophil actin deficiency, neutrophil
membrane GP-180 deficiency, metabolic storage diseases,
mucopolysaccharidoses, mucolipidoses, miscellaneous disorders
involving immune mechanisms, Wiskott-Aldrich Syndrome, alpha
lantirypsin deficiency, etc.
[0183] Diseases or pathologies include neurodegenerative diseases,
hepatodegenerative diseases, nephrodegenerative disease, spinal
cord injury, head trauma or surgery, viral infections that result
in tissue, organ, or gland degeneration, and the like. Such
neurodegenerative diseases include but are 10 not limited to, AIDS
dementia complex; demyeliriating diseases, such as multiple
sclerosis and acute transferase myelitis; extrapyramidal and
cerebellar disorders, such as lesions of the ecorticospinal system;
disorders of the basal ganglia or cerebellar disorders;
hyperkinetic movement disorders, such as Huntington's Chorea and
senile chorea; drug-induced movement disorders, such as those
induced by drugs that block CNS dopamine receptors; hypokinetic
movement disorders, such as Parkinson's disease; progressive
supra-nucleo palsy; structural lesions of the cerebellum;
spinocerebellar degenerations, such as spinal ataxia, Friedreich's
ataxia, cerebellar cortical degenerations, multiple systems
degenerations (Mencel, Dejerine Thomas, Shi-Drager, and
Machado-Joseph), systermioc disorders, such as Rufsum's disease,
abetalipoprotemia, ataxia, telangiectasia; and mitochondrial
multisystem disorder; demyelinating core disorders, such as
multiple sclerosis, acute transverse myelitis; and disorders of the
motor unit, such as neurogenic muscular atrophies (anterior horn
cell degeneration, such as amyotrophic lateral sclerosis, infantile
spinal muscular atrophy and juvenile spinal muscular atrophy);
Alzheimer's disease; Down's Syndrome in middle age; Diffuse Lewy
body disease; Senile Demetia of Lewy body type; Parkinson's
Disease, Wernicke-Korsakoff syndrome; chronic alcoholism;
Creutzfeldt-Jakob disease; Subacute sclerosing panencephalitis
hallefforden-Spatz disease; and Dementia pugilistica. See, e.g.,
Berkow et. al., (eds.) (1987), The Merck Manual, (15') ed.), Merck
and Co., Rahway, N.J.
Industrial Farming Uses
[0184] The present invention provides viable porcine for purposes
of farming applications in which one or both alleles of the
CMP-Neu5Ac hydroxylase gene have been inactivated. Inactivation of
one or both alleles of the CMP-Neu5Ac hydroxylase gene can reduce
the susceptibility of porcine animals to zoonotic diseases and
infections in pigs such as, for example, E. coli, pig rotavirus,
and pig transmissible gastroenteritis coronavirus, and any other
zoonotic or entertoxigenic organism that utilizes Neu5Gc in a host
animal. The reduction in disease susceptibility allows greater
economic realization of farming operations due to the ability to
harvest more healthy animals, and the reduction of animal death due
to enterotoxigenic organisms.
[0185] The following examples are offered by way of illustration
and not by way of limitation.
EXAMPLES
[0186] Isolation of Nucleic Acids Combination strategy of PCR-based
methods was employed to identify the porcine CMP-Neu5Ac hydroxylase
gene. Such PCR methods are well known in the art and described, for
example, in PCR Technology, H. A. Erlich, ed., Stockton Press,
London, 1989; PCR Protocols: A Guide to Methods and Applications,
M. A. Innis, D. H. Gelfand, J. J. Sninsky, and T. J. White, eds.,
Academic Pres, Inc., New York, 1990.
[0187] Total RNA was extracted from an adult porcine (Great
Yorkshire) spleen using Trizol reagent (Gibco, Grand Island, N.Y.).
After treatment with Dnase I (Ambion, Inc., Austin, Tex.), poly
A.sup.+ RNA was separated using the Dynabeads mRNA Purification Kit
(Dynal, Oslo, Norway). To identify the 5'- or 3'-end of porcine
CMP-Neu5Ac Hydroxylase gene, 5'- or 3'-RACE (rapid amplification of
cDNA ends) procedures were performed using Marathon.TM. cDNA
Amplification kit (Clontech). To identify exon-intron boundaries,
or 5'- or 3'-flanking region of the transcripts, porcine
GenomeWalker.TM. libraries were constructed using Universal
GenomeWalkeer.TM. Library kit (Clontech). Gene-specific and nested
primer pairs were designed from the partial cDNA sequence provided
by GenBank Accession #A59058.
[0188] Determination of cDNA and Genomic CMP-Neu5Ac Hydroxylase
Sequence
[0189] 5'- or 3'-RACE analysis: To identify the 5' and 3' ends of
porcine CMP-Neu5Ac hydroxylase gene transcripts, 5'-and 3'-RACE
procedures were performed using the Marathon cDNA Amplification Kit
(Clontech) with poly A+ RNA isolated from adult porcine spleen as a
template. First strand cDNA synthesis from 1 ug of poly A+ RNA was
accomplished using 20 U of AMV-RT and 1 pmol of the supplied cDNA
Synthesis Primer by incubating at 48.degree. C. for 2 hours. Second
strand cDNA synthesis involved incubating th entire first strand
reaction with a supplied enzyme cocktail composed of Rnase H, E.
coli DNA polymerase I, and E. coli DNA polymerase I, and E. coli
DNA ligase at 16.degree. C. for 1.5 hr. After blunting of the
double stranded cDNA ends by T4 DNA polymerase, the supplied
Marathon cDNA Adapters were ligated to an aliquot of purified,
double-stranded cDNA. Dilution of ht eadapter-ligated product in 10
mM ticme-KOH/0.1 mM EDTA buffer provided with the kit readied the
cDNA for PCR amplification.
[0190] To obtain the 5'- and 3'- most sequences of the porcine
CMP-Neu5Ac hydroxylase gene transcripts, provided Marathon cDNA
Amplification primer sets were paired with gene-specific and nested
gene-specific primers based on the sequence provided by GenBank
accession number A59058. These primer sets are provided for in
Table 13. By this method, oligonucleotide primers based on the
sequence contained in Genbank accession number A59058 are oriented
in the 3' and 5' directions and are used to generate overlapping
PCR fragments. These overlapping 3' and 5' products are combined to
produce an intact full-length cDNA. This method is described, for
example, in Innis, et al., supra; and Frohman et al., Proc. Natl.
Acad. Sci., 85:8998, 1988, and further described, for example, in
U.S. Pat. No. 4,683,195.
[0191] Genome Walking analysis: To identify exon-intron boundaries,
or 5'- or 3'-flanking region of the porcine CMP-Neu5Ac hydroxylase
transcripts, porcine GenomeWalker.TM. libraries were constructed
using the Universal GenomeWalker.TM. Library Kit (Clontech, Palo
Alto, Calif.).
[0192] Briefly, five aliquots of porcine genomic DNA were
separately digested with a single blunt-cutting restriction
endonuclease (DraI, EcoRV, PvuII, ScaI, or StuI). After
phenol-chloroform extraction, ethanol precipitation, and
resuspension of the restricted fragments, a portion of each
digested aliquot was used in separate ligation reaction with the
GenomeWalker adapters provided with the kit. This process created
five libraries for use in the PCR based cloning strategy. Primer
pairs identified in Table 13 were used in a genome walking
strategy. Either eLON-Gase or TaKaRa LA Taq (Takara Shuzo Co.,
Ltd., Shiga, Japan) enzyme was used for PCR in all GenomeWalker
experiments as well as for direct long PCR of genomic DNA. The
thermal cycling conditions recommended by the manufacturer were
employed in all GenomeWalker-PCR experiments on a Perkin Elmer Gene
Amp System 9600 or 9700 thermocycler.
16TABLE 13 Primers Used in PCR Strategies Primer Set PCR Strategy
Sequence XA 3'-RACE/Genome 5'-CATGGACCTCAAGCTGGGGGACAAG Walking
A-3' XB 3'-RACE/Genome 5'-GTGTTCGACCCTTGGTTAATCGGTCC Walking TG-3'
XM 5'-RACE/Genome 5'-CAGGACCGATTAACCAAGGGTCGAAC Walking AC-3' XN
5'-RACE/Genome 5'-TCTTGTCCCCCAGCTTGAGGTCCAT Walking G-3'
[0193] Subcloning and sequencing of amplified products: PCR
products amplified from genomic DNA, GeneWalker-PCR (Clontech), and
5'-3'-RACE wre gel-purified using the Qiagen Gel Extraction Kit
(Qiagen, Valencia, Calif.), if necessary, then subcloned into the
pCRII vector provided with the Original TA Cloning Kit (Invitrogen,
Carlsbad, Calif.). Plasmid DNA minipreps of pCRII-ligated inserts
were prepared with the QIAprep Spin Miniprep Kit (Qiagen) as
directed. Automated fluorescent sequencing of cloned inserts was
performed using an ABI 377 Automated Sequence Analyzer (Applied
Biosystems, Inc., Foster City, Calif.) with either the dRhodamine
or BigDye Terminator Cycle Sequencing Kits (Applied Biosystems)
primed with T7 and SP6 promoter primers or primers designed from
internal insert sequences.
[0194] Primer Synthesis: All oligonucleotides used as primers in
the various PCR-based methods were synthesized on an ABI 394 DNA
Synthesizer (Applied Biosystems, Inc., Foster City Calif.) using
solid phase synthesis and phosphoramidite nucleoside chemistry,
unless otherwise stated.
[0195] Analysis of Transcription Factor Binding Sites
[0196] Analysis of possible transcription factors binding sites
were performed using 228 bp of exon 1 sequence and 601 bp upstream
of exon 1. The sequences were screened using "Matlnspector"
software available in www.genomatix.de. The sequences contain
binding sites for the following transcription factors: MZFI, ETSF,
SF1, CMYB, MEF2, NMP4, BRN2, API, GATI, SATBI, ATF, USF, WHN, ZF5,
NF.kappa.B, MOK2, NFY, MYCMAX, ZF5. See FIG. 4.
[0197] Construction of Porcine CMP-Neu5Ac Hydroxylase Homologous
Recombination Targeting Vectors
[0198] CMP-Neu5Ac hydroxylase knock-out target vector: A vector
targeting Exon 6 of the porcine CMP-Neu5Ac Hydroxylase gene for
knockout can be constructed. In a first step, a portion of Intron 6
is amplified by PCR for use as a 3'-arm of the targeting vector
utilizing primers such as pDH3 (5'-CTCCTGGAAGCTTCTGTCAAGACGAAC-3')
and pDH4 (5'-GCCTGATACACAGTGCTGT- GCAATGGT-3') (see FIG. 5). The
amplified PCR product of approximately 3.7 kb can be inserted into
the pCRII vector after restriction enzyme digestion utilizing EcoRI
and ApaI. See FIG. 6.
[0199] Following the insertion of the 3'-arm, a portion of Intron 5
can be amplified by PCR for use as a 5'-arm in the targeting vector
utilizing primers such as pDH1
(5'-ACCACCCAAGTCTGGAATCTTCTTACACT-3') and pDH2
(5'-GACTCTCATACAAAAGCTAAGCTGGGTAAG-3') (see FIG. 5). Following this
initial amplification, successive PCR amplifications can be
performed to introduce an EcoNI restriction site into the 3'
portion of the 5'-arm utilizing primers such as pDH1 in conjunction
with primers such as pDH2a (5'-GACTCTCATACAAAACCTAAGCTGGGTAAG-3'),
pDH2b (5'-GACTCTCATACAAAACCTAGGCT- GGGTAAG-3'), and pDH2c
(5'-GACTCTCATACAAAACCTAGGCTAGGTAAG-3'), respectively (see FIG. 5).
The amplified PCR product of approximately 2.6 kb containing the
engineered EcoNI site can be restriction enzyme digested using ApaI
and EcoNI, and inserted into the pCRII vector containing the
previously inserted 3'-arm (See FIG. 7), generating a targeting
vector (pDH.DELTA.ex6) containing an approximate 6.3 kb porcine
CMP-Neu5Ac hydroxylase targeting sequence (see FIG. 8).
[0200] EGFP knock-in target vector: pDH.DELTA.ex6 can be further
modified by an in-frame insertion of an enhanced green fluorescent
protein sequence at the terminal 3' end of Exon 6 of the porcine
CMP-Neu5Ac hydroxylase gene. In a first step, a portion of Intron 5
and a portion of Exon 6 of the porcine CMP-Neu5Ac hydroxylase gene
can be amplified by PCR utilizing primers such as pDH5
(5'-CCTTATACTGGCCCCAATTGGATCTTAC-3') and pDH6
(5'-CCTTATACTGGCCCCAATTGGATCTTAC-3') (see FIG. 9), and inserted
into a vector (pIRES-EGFP) containing the EGFP and a poly A tail
following restriction enzyme digestion with MunI and EcoRv.
Following insertion, PCR amplification can be performed on the
pIRES-EGFP vector containing the insertion utilizing primer such as
pDH7 (5'-CTTACCTAGCCTAGGTTTTGTATGA- GAGTC-3') and pDH8
(5'-GACAAACCACAATTGGAATGCACTCGAG-3') (see FIG. 9). The PCR
amplified product can be restriction enzyme digested using EcoNI
and MunI and inserted into the previously constructed pDH.DELTA.ex6
targeting vector (see FIG. 10). The resultant targeting vector
(pDH.DELTA.ex6-EGFP) is illustrated in FIG. 11.
[0201] Production of Porcine CMP-Neu5Ac Hydroxylase Deficient Fetal
Fibroblast Cells
[0202] Fetal fibroblast cells are isolated from 10 fetuses of the
same pregnancy at day 33 of gestation. After removing the head and
viscera, fetuses are washed with Hanks' balanced salt solution
(HBSS; Gibco-BRL, 1 5 Rockville, Md.), placed in 20 ml of HBSS, and
diced with small surgical scissors. The tissue is pelleted and
resuspended in 50-ml tubes with 40 ml of DMEM and 100 U/ml
collagenase (Gibco-BRL) per fetus. Tubes are incubated for 40 min
in a shaking water bath at 37C. The digested tissue is allowed to
settle for 3-4 min and the cell-rich supernatant is transferred to
a new 50-ml tube and pelleted. The cells are then resuspended in 40
ml of DMEM containing 10% fetal calf serum (FCS), 1X nonessential
amino acids, 1 mM sodium pyruvate and 2 ng/ml bFGF, and seeded into
10 cm. dishes. For transfections, 10 .mu.g of linearized
pDH.DELTA.ex6EGFP vector is introduced into 2 million cells using
lipofectamine 2000 (Carlsbad, Calif.) following manufacturer's
guidelines. Forty-eight hours after transfection, the transfected
cells are seeded into 48-well plates at a density of 2,000 cells
per well and grown to confluence. Following confluence, cells are
sorted via Fluorescent Activated Cell Sorting (FACS) (FACSCalibur,
Becton Dickenson, San Jose, Calif.), wherein only cells having
undergone homologous recombination and expressing the EGFP are
selected (see, for example, FIG. 13).
[0203] Selected cells are then reseeded, and grown to confluency.
Once confluency is reached, several small aliquots are frozen back
for future use, and the remainder are utilized for PCR and Southern
Blot verification of homologous recombination. The putative
targeted clones can be screened by PCR across the Exon 6/EGFP
insert utilizing a primer complimentary to the EGFP sequence and a
primer complimentary to a sequence outside the vector as the
antisense primer. The PCR products can be analyzed by Southern
Blotting using an EGFP probe to identify the positive clones by the
presence of the expected band from the targeted allele.
[0204] Generation of Cloned Pigs Using Heterologous CMP-Neu5Ac
Hydroxylase Deficient Fetal Fibroblasts as Nuclear Donors
[0205] Preparation of cells for Nuclear Transfer: Donor cells are
genetically manipulated to produce cells heterozygous for porcine
CMP-Neu5Ac hydroxylase as described generally above. Nuclear
transfer can be performed by methods that are well known in the art
(see, e.g., Dai et al., Nature Biotechnology 20: 251255, 2002; and
Polejaeva et al., Nature 407:86-90, 2000), using EGFP selected
porcine fibroblasts as nuclear donors that are produced as
described in detail hereinabove.
[0206] Oocytes can be isolated from synchronized super ovulated
sexually mature Large-White X Landacre outcross gilts as described,
for example, in I. Polejaeva et al. Nature 407: 505 (2000). Donor
cells are synchronized in presumptive G0/G1 by serum starvation
(0.5%) between 24 to 120 hours. Oocytes enucleation, nuclear
transfer, electrofusion, and electroactivation can be performed as
essentially described in, for example, A. C. Boquest et al., Biol.
Reproduction 68: 1283 (2002). Reconstructed embryos can be cultured
overnight and can be transferred to the oviducts of asynchronous
(-1 day) recipients. Pregnancies can be confirmed and monitored by
real-time ultrasound.
[0207] Breeding of heterozygous CMP-Neu5Ac hydroxylase single
knockout (SKO) male and female pigs can be performed to establish a
miniherd of double knockout (DKO) pigs.
[0208] Verification of CMP-Neu5Ac Hydroxylase Deficient Pigs
[0209] Following breeding of the single knockout male and female
pigs, verification of double knockout pigs is performed.
Fibroblasts from the offspring are incubated with 1 .mu.g of
anti-N-glycolyl GM2 monoclonal antibody MK2-34 (Seikagaku Kogyo,
JP) on ice for 30 minutes. FITC conjugated goat-anti-mouse IgG is
added to the cells and antibody binding indicating the presence or
absence of Neu5GC, and thus, an indication of the presence or
absence of active CMP-Neu5Ac hydroxylase, is detected by flow
cytometry (FACSCalibur, Becton Dickenson, San Jose, Calif.).
Sequence CWU 1
1
65 1 2728 DNA Porcine 1 cccgacgtcc tggcagcgcc caggcactgt tattggtgcc
tcctgtgtcc acgcgcttcc 60 cggccaggca gccctggcgg atcctatttt
ctgttccccc gattctggta cctctccctc 120 ccgccctcgg tgcgcagccg
tcctcctgca gtgcctgctc ctccaggggc gaaaccgatc 180 agggatcagg
ccacccgcct cctgaacatc cctccttagt tcccacagtc taatgccttg 240
tggaagcaaa tgagccacag aagctgaagg aaaaaccacc attctttctt aatacctgga
300 gagaggcaac gacagactat gagcagcatc gaacaaacga cggagatcct
gttgtgcctc 360 tcacctgccg aagctgccaa tctcaaggaa ggaatcaatt
ttgttcgaaa taagagcact 420 ggcaaggatt acatcttatt taagaataag
agccgcctga aggcatgtaa gaacatgtgc 480 aagcaccaag gaggcctctt
cattaaagac attgaggatc taaatggaag gtctgttaaa 540 tgcacaaaac
acaactggaa gttagatgta agcagcatga agtatatcaa tcctcctgga 600
agcttctgtc aagacgaact ggttgtagaa aaggatgaag aaaatggagt tttgcttcta
660 gaactaaatc ctcctaaccc gtgggattca gaacccagat ctcctgaaga
tttggcattt 720 ggggaagtgc agatcacgta ccttactcac gcctgcatgg
acctcaagct gggggacaag 780 agaatggtgt tcgacccttg gttaatcggt
cctgcttttg cgcgaggatg gtggttacta 840 cacgagcctc catctgattg
gctggagagg ctgagccgcg cagacttaat ttacatcagt 900 cacatgcact
cagaccacct gagttaccca acactgaaga agcttgctga gagaagacca 960
gatgttccca tttatgttgg caacacggaa agacctgtat tttggaatct gaatcagagt
1020 ggcgtccagt tgactaatat caatgtagtg ccatttggaa tatggcagca
ggtagacaaa 1080 aatcttcgat tcatgatctt gatggatggc gttcatcctg
agatggacac ttgcattatt 1140 gtggaataca aaggtcataa aatactcaat
acagtggatt gcaccagacc caatggagga 1200 aggctgccta tgaaggttgc
attaatgatg agtgattttg ctggaggagc ttcaggcttt 1260 ccaatgactt
tcagtggtgg aaaatttact gaggaatgga aagcccaatt cattaaaaca 1320
gaaaggaaga aactcctgaa ctacaaggct cggctggtga aggacctaca acccagaatt
1380 tactgcccct ttcctgggta tttcgtggaa tcccacccag cagacaagta
tattaaggaa 1440 acaaacatca aaaatgaccc aaatgaactc aacaatctta
tcaagaagaa ttctgaggtg 1500 gtaacctgga ccccaagacc tggagccact
cttgatctgg gtaggatgct aaaggaccca 1560 acagacagca agggcatcgt
agagcctcca gaagggacta agatttacaa ggattcctgg 1620 gattttggcc
catatttgaa tatcttgaat gctgctatag gagatgaaat atttcgtcac 1680
tcatcctgga taaaagaata cttcacttgg gctggattta aggattataa cctggtggtc
1740 aggatgattg agacagatga ggacttcagc cctttgcctg gaggatatga
ctatttggtt 1800 gactttctgg atttatcctt tccaaaagaa agaccaagcc
gggaacatcc atatgaggaa 1860 attcggagcc gggttgatgt catcagacac
gtggtaaaga atggtctgct ctgggatgac 1920 ttgtacatag gattccaaac
ccggcttcag cgggatcctg atatatacca tcatctgttt 1980 tggaatcatt
ttcaaataaa actccccctc acaccacctg actggaagtc cttcctgatg 2040
tgctctgggt agagaggacc tgagctgtcc caggggtgcc caacaacatg aaaaaatcaa
2100 gaatttattg ctgctacgtc aaagcttata ccagagatta tgccttatag
acattagcaa 2160 tggataatta tatgttgcac ttgtgaaatg tgcacatatc
ctgtttatga atcaccacat 2220 agccagatta tcaatatttt acttatttcg
taaaaaatcc acaattttcc ataacagaat 2280 caacgtgtgc aataggaaca
agattgctat ggaaaacgag ggtaacagga ggagatatta 2340 atccaagcat
agaagaaata gacaaatgag gggccataag gggaatatag ggaagagaaa 2400
aaaattaaga tggaatttta aaaggagaat gtaaaaaata gatatttgtt ccttaatagg
2460 ttgattcctc aaatagagcc catgaatata atcaaatagg aagggttcat
gactgttttc 2520 aatttttcaa aaagctttgt tgaaatcata gacttgcaaa
acaaggctgt agaggccacc 2580 ctaaaatgga aaatttcact gggactgaaa
ttattttgat tcaatgacaa aatttgttat 2640 ttactgcgga ttataaactc
taacaaatag cgatctcttt gcttcataaa aacataaaca 2700 ctagctagta
ataaaatgag ttctgcag 2728 2 577 PRT Porcine 2 Met Ser Ser Ile Glu
Gln Thr Thr Glu Ile Leu Leu Cys Leu Ser Pro 1 5 10 15 Ala Glu Ala
Ala Asn Leu Lys Glu Gly Ile Asn Phe Val Arg Asn Lys 20 25 30 Ser
Thr Gly Lys Asp Tyr Ile Leu Phe Lys Asn Lys Ser Arg Leu Lys 35 40
45 Ala Cys Lys Asn Met Cys Lys His Gln Gly Gly Leu Phe Ile Lys Asp
50 55 60 Ile Glu Asp Leu Asn Gly Arg Ser Val Lys Cys Thr Lys His
Asn Trp 65 70 75 80 Lys Leu Asp Val Ser Ser Met Lys Tyr Ile Asn Pro
Pro Gly Ser Phe 85 90 95 Cys Gln Asp Glu Leu Val Val Glu Lys Asp
Glu Glu Asn Gly Val Leu 100 105 110 Leu Leu Glu Leu Asn Pro Pro Asn
Pro Trp Asp Ser Glu Pro Arg Ser 115 120 125 Pro Glu Asp Leu Ala Phe
Gly Glu Val Gln Ile Thr Tyr Leu Thr His 130 135 140 Ala Cys Met Asp
Leu Lys Leu Gly Asp Lys Arg Met Val Phe Asp Pro 145 150 155 160 Trp
Leu Ile Gly Pro Ala Phe Ala Arg Gly Trp Trp Leu Leu His Glu 165 170
175 Pro Pro Ser Asp Trp Leu Glu Arg Leu Ser Arg Ala Asp Leu Ile Tyr
180 185 190 Ile Ser His Met His Ser Asp His Leu Ser Tyr Pro Thr Leu
Lys Lys 195 200 205 Leu Ala Glu Arg Arg Pro Asp Val Pro Ile Tyr Val
Gly Asn Thr Glu 210 215 220 Arg Pro Val Phe Trp Asn Leu Asn Gln Ser
Gly Val Gln Leu Thr Asn 225 230 235 240 Ile Asn Val Val Pro Phe Gly
Ile Trp Gln Gln Val Asp Lys Asn Leu 245 250 255 Arg Phe Met Ile Leu
Met Asp Gly Val His Pro Glu Met Asp Thr Cys 260 265 270 Ile Ile Val
Glu Tyr Lys Gly His Lys Ile Leu Asn Thr Val Asp Cys 275 280 285 Thr
Arg Pro Asn Gly Gly Arg Leu Pro Met Lys Val Ala Leu Met Met 290 295
300 Ser Asp Phe Ala Gly Gly Ala Ser Gly Phe Pro Met Thr Phe Ser Gly
305 310 315 320 Gly Lys Phe Thr Glu Glu Trp Lys Ala Gln Phe Ile Lys
Thr Glu Arg 325 330 335 Lys Lys Leu Leu Asn Tyr Lys Ala Arg Leu Val
Lys Asp Leu Gln Pro 340 345 350 Arg Ile Tyr Cys Pro Phe Pro Gly Tyr
Phe Val Glu Ser His Pro Ala 355 360 365 Asp Lys Tyr Ile Lys Glu Thr
Asn Ile Lys Asn Asp Pro Asn Glu Leu 370 375 380 Asn Asn Leu Ile Lys
Lys Asn Ser Glu Val Val Thr Trp Thr Pro Arg 385 390 395 400 Pro Gly
Ala Thr Leu Asp Leu Gly Arg Met Leu Lys Asp Pro Thr Asp 405 410 415
Ser Lys Gly Ile Val Glu Pro Pro Glu Gly Thr Lys Ile Tyr Lys Asp 420
425 430 Ser Trp Asp Phe Gly Pro Tyr Leu Asn Ile Leu Asn Ala Ala Ile
Gly 435 440 445 Asp Glu Ile Phe Arg His Ser Ser Trp Ile Lys Glu Tyr
Phe Thr Trp 450 455 460 Ala Gly Phe Lys Asp Tyr Asn Leu Val Val Arg
Met Ile Glu Thr Asp 465 470 475 480 Glu Asp Phe Ser Pro Leu Pro Gly
Gly Tyr Asp Tyr Leu Val Asp Phe 485 490 495 Leu Asp Leu Ser Phe Pro
Lys Glu Arg Pro Ser Arg Glu His Pro Tyr 500 505 510 Glu Glu Ile Arg
Ser Arg Val Asp Val Ile Arg His Val Val Lys Asn 515 520 525 Gly Leu
Leu Trp Asp Asp Leu Tyr Ile Gly Phe Gln Thr Arg Leu Gln 530 535 540
Arg Asp Pro Asp Ile Tyr His His Leu Phe Trp Asn His Phe Gln Ile 545
550 555 560 Lys Leu Pro Leu Thr Pro Pro Asp Trp Lys Ser Phe Leu Met
Cys Ser 565 570 575 Gly 3 2467 DNA Porcine 3 cccgacgtcc tggcagcgcc
caggcactgt tattggtgcc tcctgtgtcc acgcgcttcc 60 cggccaggca
gccctggcgg atcctatttt ctgttccccc gattctggta cctctccctc 120
ccgccctcgg tgcgcagccg tcctcctgca gtgcctgctc ctccaggggc gaaaccgatc
180 agggatcagg ccacccgcct cctgaacatc cctccttagt tcccacagtc
taatgccttg 240 tggaagcaaa tgagccacag aagctgaagg aaaaaccacc
attctttctt aatacctgga 300 gagaggcaac gacagactat gagcagcatc
gaacaaacga cggagatcct gttgtgcctc 360 tcacctgccg aagctgccaa
tctcaaggaa ggaatcaatt ttgttcgaaa taagagcact 420 ggcaaggatt
acatcttatt taagaataag agccgcctga aggcatgtaa gaacatgtgc 480
aagcaccaag gaggcctctt cattaaagac attgaggatc taaatggaag gtctgttaaa
540 tgcacaaaac acaactggaa gttagatgta agcagcatga agtatatcaa
tcctcctgga 600 agcttctgtc aagacgaact ggttgtagaa aaggatgaag
aaaatggagt tttgcttcta 660 gaactaaatc ctcctaaccc gtgggattca
gaacccagat ctcctgaaga tttggcattt 720 ggggaagtgc agatcacgta
ccttactcac gcctgcatgg acctcaagct gggggacaag 780 agaatggtgt
tcgacccttg gttaatcggt cctgcttttg cgcgaggatg gtggttacta 840
cacgagcctc catctgattg gctggagagg ctgagccgcg cagacttaat ttacatcagt
900 cacatgcact cagaccacct gagttaccca acactgaaga agcttgctga
gagaagacca 960 gatgttccca tttatgttgg caacacggaa agacctgtat
tttggaatct gaatcagagt 1020 ggcgtccagt tgactaatat caatgtagtg
ccatttggaa tatggcagca ggtagacaaa 1080 aatcttcgat tcatgatctt
gatggatggc gttcatcctg agatggacac ttgcattatt 1140 gtggaataca
aaggtcataa aatactcaat acagtggatt gcaccagacc caatggagga 1200
aggctgccta tgaaggttgc attaatgatg agtgattttg ctggaggagc ttcaggcttt
1260 ccaatgactt tcagtggtgg aaaatttact gaggaatgga aagcccaatt
cattaaaaca 1320 gaaaggaaga aactcctgaa ctacaaggct cggctggtga
aggacctaca acccagaatt 1380 tactgcccct ttcctgggta tttcgtggaa
tcccacccag cagacaagta tattaaggaa 1440 acaaacatca aaaatgaccc
aaatgaactc aacaatctta tcaagaagaa ttctgaggtg 1500 gtaacctgga
ccccaagacc tggagccact cttgatctgg gtaggatgct aaaggaccca 1560
acagacagat cctgtgtcag gagttgggat tctttgaaga ttcggagccg ggttgatgtc
1620 atcagacacg tggtaaagaa tggtctgctc tgggatgact tgtacatagg
attccaaacc 1680 cggcttcagc gggatcctga tatataccat catctgtttt
ggaatcattt tcaaataaaa 1740 ctccccctca caccacctga ctggaagtcc
ttcctgatgt gctctgggta gagaggacct 1800 gagctgtccc aggggtgccc
aacaacatga aaaaatcaag aatttattgc tgctacgtca 1860 aagcttatac
cagagattat gccttataga cattagcaat ggataattat atgttgcact 1920
tgtgaaatgt gcacatatcc tgtttatgaa tcaccacata gccagattat caatatttta
1980 cttatttcgt aaaaaatcca caattttcca taacagaatc aacgtgtgca
ataggaacaa 2040 gattgctatg gaaaacgagg gtaacaggag gagatattaa
tccaagcata gaagaaatag 2100 acaaatgagg ggccataagg ggaatatagg
gaagagaaaa aaattaagat ggaattttaa 2160 aaggagaatg taaaaaatag
atatttgttc cttaataggt tgattcctca aatagagccc 2220 atgaatataa
tcaaatagga agggttcatg actgttttca atttttcaaa aagctttgtt 2280
gaaatcatag acttgcaaaa caaggctgta gaggccaccc taaaatggaa aatttcactg
2340 ggactgaaat tattttgatt caatgacaaa atttgttatt tactgcggat
tataaactct 2400 aacaaatagc gatctctttg cttcataaaa acataaacac
tagctagtaa taaaatgagt 2460 tctgcag 2467 4 490 PRT Porcine 4 Met Ser
Ser Ile Glu Gln Thr Thr Glu Ile Leu Leu Cys Leu Ser Pro 1 5 10 15
Ala Glu Ala Ala Asn Leu Lys Glu Gly Ile Asn Phe Val Arg Asn Lys 20
25 30 Ser Thr Gly Lys Asp Tyr Ile Leu Phe Lys Asn Lys Ser Arg Leu
Lys 35 40 45 Ala Cys Lys Asn Met Cys Lys His Gln Gly Gly Leu Phe
Ile Lys Asp 50 55 60 Ile Glu Asp Leu Asn Gly Arg Ser Val Lys Cys
Thr Lys His Asn Trp 65 70 75 80 Lys Leu Asp Val Ser Ser Met Lys Tyr
Ile Asn Pro Pro Gly Ser Phe 85 90 95 Cys Gln Asp Glu Leu Val Val
Glu Lys Asp Glu Glu Asn Gly Val Leu 100 105 110 Leu Leu Glu Leu Asn
Pro Pro Asn Pro Trp Asp Ser Glu Pro Arg Ser 115 120 125 Pro Glu Asp
Leu Ala Phe Gly Glu Val Gln Ile Thr Tyr Leu Thr His 130 135 140 Ala
Cys Met Asp Leu Lys Leu Gly Asp Lys Arg Met Val Phe Asp Pro 145 150
155 160 Trp Leu Ile Gly Pro Ala Phe Ala Arg Gly Trp Trp Leu Leu His
Glu 165 170 175 Pro Pro Ser Asp Trp Leu Glu Arg Leu Ser Arg Ala Asp
Leu Ile Tyr 180 185 190 Ile Ser His Met His Ser Asp His Leu Ser Tyr
Pro Thr Leu Lys Lys 195 200 205 Leu Ala Glu Arg Arg Pro Asp Val Pro
Ile Tyr Val Gly Asn Thr Glu 210 215 220 Arg Pro Val Phe Trp Asn Leu
Asn Gln Ser Gly Val Gln Leu Thr Asn 225 230 235 240 Ile Asn Val Val
Pro Phe Gly Ile Trp Gln Gln Val Asp Lys Asn Leu 245 250 255 Arg Phe
Met Ile Leu Met Asp Gly Val His Pro Glu Met Asp Thr Cys 260 265 270
Ile Ile Val Glu Tyr Lys Gly His Lys Ile Leu Asn Thr Val Asp Cys 275
280 285 Thr Arg Pro Asn Gly Gly Arg Leu Pro Met Lys Val Ala Leu Met
Met 290 295 300 Ser Asp Phe Ala Gly Gly Ala Ser Gly Phe Pro Met Thr
Phe Ser Gly 305 310 315 320 Gly Lys Phe Thr Glu Glu Trp Lys Ala Gln
Phe Ile Lys Thr Glu Arg 325 330 335 Lys Lys Leu Leu Asn Tyr Lys Ala
Arg Leu Val Lys Asp Leu Gln Pro 340 345 350 Arg Ile Tyr Cys Pro Phe
Pro Gly Tyr Phe Val Glu Ser His Pro Ala 355 360 365 Asp Lys Tyr Ile
Lys Glu Thr Asn Ile Lys Asn Asp Pro Asn Glu Leu 370 375 380 Asn Asn
Leu Ile Lys Lys Asn Ser Glu Val Val Thr Trp Thr Pro Arg 385 390 395
400 Pro Gly Ala Thr Leu Asp Leu Gly Arg Met Leu Lys Asp Pro Thr Asp
405 410 415 Arg Ser Cys Val Arg Ser Trp Asp Ser Leu Lys Ile Arg Ser
Arg Val 420 425 430 Asp Val Ile Arg His Val Val Lys Asn Gly Leu Leu
Trp Asp Asp Leu 435 440 445 Tyr Ile Gly Phe Gln Thr Arg Leu Gln Arg
Asp Pro Asp Ile Tyr His 450 455 460 His Leu Phe Trp Asn His Phe Gln
Ile Lys Leu Pro Leu Thr Pro Pro 465 470 475 480 Asp Trp Lys Ser Phe
Leu Met Cys Ser Gly 485 490 5 1624 DNA Porcine 5 cccgacgtcc
tggcagcgcc caggcactgt tattggtgcc tcctgtgtcc acgcgcttcc 60
cggccaggca gccctggcgg atcctatttt ctgttccccc gattctggta cctctccctc
120 ccgccctcgg tgcgcagccg tcctcctgca gtgcctgctc ctccaggggc
gaaaccgatc 180 agggatcagg ccacccgcct cctgaacatc cctccttagt
tcccacagtc taatgccttg 240 tggaagcaaa tgagccacag aagctgaagg
aaaaaccacc attctttctt aatacctgga 300 gagaggcaac gacagactat
gagcagcatc gaacaaacga cggagatcct gttgtgcctc 360 tcacctgccg
aagctgccaa tctcaaggaa ggaatcaatt ttgttcgaaa taagagcact 420
ggcaaggatt acatcttatt taagaataag agccgcctga aggcatgtaa gaacatgtgc
480 aagcaccaag gaggcctctt cattaaagac attgaggatc taaatggaag
gtctgttaaa 540 tgcacaaaac acaactggaa gttagatgta agcagcatga
agtatatcaa tcctcctgga 600 agcttctgtc aagacgaact ggttgtagaa
aaggatgaag aaaatggagt tttgcttcta 660 gaactaaatc ctcctaaccc
gtgggattca gaacccagat ctcctgaaga tttggcattt 720 ggggaagtgc
agatcacgta ccttactcac gcctgcatgg acctcaagct gggggacaag 780
agaatggtgt tcgacccttg gttaatcggt cctgcttttg cgcgaggatg gtggttacta
840 cacgagcctc catctgattg gctggagagg ctgagccgcg cagacttaat
ttacatcagt 900 cacatgcact cagaccacct gagttaccca acactgaaga
agcttgctga gagaagacca 960 gatgttccca tttatgttgg caacacggaa
agacctgtat tttggaatct gaatcagagt 1020 ggcgtccagt tgactaatat
caatgtagtg ccatttggaa tatggcagca ggtagacaaa 1080 aatcttcgat
tcatgatctt gatggatggc gttcatcctg agatggacac ttgcattatt 1140
gtggaataca aaggtcataa aatactcaat acagtggatt gcaccagacc caatggagga
1200 aggctgccta tgaaggttgc attaatgatg agtgattttg ctggaggagc
ttcaggcttt 1260 ccaatgactt tcagtggtgg aaaatttact gaggaatgga
aagcccaatt cattaaaaca 1320 gaaaggaaga aactcctgaa ctacaaggct
cggctggtga aggacctaca acccagaatt 1380 tactgcccct ttcctgggta
tttcgtggaa tcccacccag cagacaagta tggctggata 1440 ttttatataa
cgtgtttacg cataagttaa tatatgctga atgagtgatt tagctgtgaa 1500
acaacatgaa atgagaaaga atgattagta ggggtctgga gcttatttta acaagcagcc
1560 tgaaaacaga aagtatgaat aaaaaaaatt aaatgcaaaa aaaaaaaaaa
aaaaaaaaaa 1620 aaaa 1624 6 383 PRT Porcine 6 Met Ser Ser Ile Glu
Gln Thr Thr Glu Ile Leu Leu Cys Leu Ser Pro 1 5 10 15 Ala Glu Ala
Ala Asn Leu Lys Glu Gly Ile Asn Phe Val Arg Asn Lys 20 25 30 Ser
Thr Gly Lys Asp Tyr Ile Leu Phe Lys Asn Lys Ser Arg Leu Lys 35 40
45 Ala Cys Lys Asn Met Cys Lys His Gln Gly Gly Leu Phe Ile Lys Asp
50 55 60 Ile Glu Asp Leu Asn Gly Arg Ser Val Lys Cys Thr Lys His
Asn Trp 65 70 75 80 Lys Leu Asp Val Ser Ser Met Lys Tyr Ile Asn Pro
Pro Gly Ser Phe 85 90 95 Cys Gln Asp Glu Leu Val Val Glu Lys Asp
Glu Glu Asn Gly Val Leu 100 105 110 Leu Leu Glu Leu Asn Pro Pro Asn
Pro Trp Asp Ser Glu Pro Arg Ser 115 120 125 Pro Glu Asp Leu Ala Phe
Gly Glu Val Gln Ile Thr Tyr Leu Thr His 130 135 140 Ala Cys Met Asp
Leu Lys Leu Gly Asp Lys Arg Met Val Phe Asp Pro 145 150 155 160 Trp
Leu Ile Gly Pro Ala Phe Ala Arg Gly Trp Trp Leu Leu His Glu 165 170
175 Pro Pro Ser Asp Trp Leu Glu Arg Leu Ser Arg Ala Asp Leu Ile Tyr
180 185 190 Ile Ser His Met His Ser Asp His Leu Ser Tyr Pro Thr Leu
Lys Lys 195 200 205 Leu Ala Glu Arg Arg Pro Asp Val Pro Ile Tyr Val
Gly Asn Thr Glu 210 215 220 Arg Pro Val Phe Trp Asn Leu Asn Gln Ser
Gly Val Gln Leu Thr Asn
225 230 235 240 Ile Asn Val Val Pro Phe Gly Ile Trp Gln Gln Val Asp
Lys Asn Leu 245 250 255 Arg Phe Met Ile Leu Met Asp Gly Val His Pro
Glu Met Asp Thr Cys 260 265 270 Ile Ile Val Glu Tyr Lys Gly His Lys
Ile Leu Asn Thr Val Asp Cys 275 280 285 Thr Arg Pro Asn Gly Gly Arg
Leu Pro Met Lys Val Ala Leu Met Met 290 295 300 Ser Asp Phe Ala Gly
Gly Ala Ser Gly Phe Pro Met Thr Phe Ser Gly 305 310 315 320 Gly Lys
Phe Thr Glu Glu Trp Lys Ala Gln Phe Ile Lys Thr Glu Arg 325 330 335
Lys Lys Leu Leu Asn Tyr Lys Ala Arg Leu Val Lys Asp Leu Gln Pro 340
345 350 Arg Ile Tyr Cys Pro Phe Pro Gly Tyr Phe Val Glu Ser His Pro
Ala 355 360 365 Asp Lys Tyr Gly Trp Ile Phe Tyr Ile Thr Cys Leu Arg
Ile Ser 370 375 380 7 2293 DNA Porcine 7 cccgacgtcc tggcagcgcc
caggcactgt tattggtgcc tcctgtgtcc acgcgcttcc 60 cggccaggca
gccctggcgg atcctatttt ctgttccccc gattctggta cctctccctc 120
ccgccctcgg tgcgcagccg tcctcctgca gtgcctgctc ctccaggggc gaaaccgatc
180 agggatcagg ccacccgcct cctgaacatc cctccttagt tcccacagtc
taatgccttg 240 tggaagcaaa tgagccacag aagctgaagg aaaaaccacc
attctttctt aatacctgga 300 gagaggcaac gacagactat gagcagcatc
gaacaaacga cggagatcct gttgtgcctc 360 tcacctgccg aagctgccaa
tctcaaggaa ggaatcaatt ttgttcgaaa taagagcact 420 ggcaaggatt
acatcttatt taagaataag agccgcctga aggcatgtaa gaacatgtgc 480
aagcaccaag gaggcctctt cattaaagac attgaggatc taaatggaag gtctgttaaa
540 tgcacaaaac acaactggaa gttagatgta agcagcatga agtatatcaa
tcctcctgga 600 agcttctgtc aagacgaact ggttgtagaa aaggatgaag
aaaatggagt tttgcttcta 660 gaactaaatc ctcctaaccc gtgggattca
gaacccagat ctcctgaaga tttggcattt 720 ggggaagtgc agatcacgta
ccttactcac gcctgcatgg acctcaagct gggggacaag 780 agaatggtgt
tcgacccttg gttaatcggt cctgcttttg cgcgaggatg gtggttacta 840
cacgagcctc catctgattg gctggagagg ctgagccgcg cagacttaat ttacatcagt
900 cacatgcact cagaccacct gagttaccca acactgaaga agcttgctga
gagaagacca 960 gatgttccca tttatgttgg caacacggaa agacctgtat
tttggaatct gaatcagagt 1020 ggcgtccagt tgactaatat caatgtagtg
ccatttggaa tatggcagca ggtagacaaa 1080 aatcttcgat tcatgatctt
gatggatggc gttcatcctg agatggacac ttgcattatt 1140 gtggaataca
aaggtcataa aatactcaat acagtggatt gcaccagacc caatggagga 1200
aggctgccta tgaaggttgc attaatgatg agtgattttg ctggaggagc ttcaggcttt
1260 ccaatgactt tcagtggtgg aaaatttact ggtaattctt tatatcaaaa
tgatgccaag 1320 gagttggcat ggcactttgc taaatgctgt gtgaatcaat
acaaagataa ttaggacatg 1380 gttcttcctc acaagaggtg tgcaatctta
ttgggaaatc atacttgcaa gtcacaaata 1440 tagactaaag tttccagctg
agaatatgct gatggagcat gaaacactaa ggagacaggg 1500 agaatctcag
gaaaaatcaa gaataatttg gatcaaatgg attcctgaca tagaacatag 1560
agctgatcag aaagagtctg acattggtaa tccaggctta agtgctcttt gtatgtggtt
1620 cagaacagag tgtgggcagc ctgaggggga tacataccct tgacctcgtg
gaaagctcat 1680 acgggggagg gatgaggcta aggaagcccc tctaaagtgt
gggattacga gaggttgggg 1740 gggtggtagg gaaaatagtg gtcaaagagt
ataaacttcc agttacaaga tgaataaatt 1800 ctaggggtat aataacagca
tggcactata gatagcatat tgtactatat actggaagtg 1860 ctgagagtag
atcttacatg ttctaaccac acacacacac acacacacac acacaccaca 1920
cacacacacc acacacacac acgtgcacac aaacagaaat ggtaattatg tgaggtgatg
1980 gcggtgttaa ctaactttat tgtggtcatc atttagccat acatgcatgt
catgaaatca 2040 ccatgttgta caccttaaag ttatgtaata ctagatgtca
gttatatctc aaagctagaa 2100 aaaatgtggg gaccaaggca gaagctcttc
tgctctgtgt ctaagggtgg ttctggggct 2160 gggatgggga ggatggttaa
gtggtatatt tttttcatac ctttgctcag tactatcatt 2220 gtaagtgttc
aatatatgtc tgcttaataa attaatgttt ttagtaaaaa aaaaaaaaaa 2280
aaaaaaaaaa aaa 2293 8 351 PRT Porcine 8 Met Ser Ser Ile Glu Gln Thr
Thr Glu Ile Leu Leu Cys Leu Ser Pro 1 5 10 15 Ala Glu Ala Ala Asn
Leu Lys Glu Gly Ile Asn Phe Val Arg Asn Lys 20 25 30 Ser Thr Gly
Lys Asp Tyr Ile Leu Phe Lys Asn Lys Ser Arg Leu Lys 35 40 45 Ala
Cys Lys Asn Met Cys Lys His Gln Gly Gly Leu Phe Ile Lys Asp 50 55
60 Ile Glu Asp Leu Asn Gly Arg Ser Val Lys Cys Thr Lys His Asn Trp
65 70 75 80 Lys Leu Asp Val Ser Ser Met Lys Tyr Ile Asn Pro Pro Gly
Ser Phe 85 90 95 Cys Gln Asp Glu Leu Val Val Glu Lys Asp Glu Glu
Asn Gly Val Leu 100 105 110 Leu Leu Glu Leu Asn Pro Pro Asn Pro Trp
Asp Ser Glu Pro Arg Ser 115 120 125 Pro Glu Asp Leu Ala Phe Gly Glu
Val Gln Ile Thr Tyr Leu Thr His 130 135 140 Ala Cys Met Asp Leu Lys
Leu Gly Asp Lys Arg Met Val Phe Asp Pro 145 150 155 160 Trp Leu Ile
Gly Pro Ala Phe Ala Arg Gly Trp Trp Leu Leu His Glu 165 170 175 Pro
Pro Ser Asp Trp Leu Glu Arg Leu Ser Arg Ala Asp Leu Ile Tyr 180 185
190 Ile Ser His Met His Ser Asp His Leu Ser Tyr Pro Thr Leu Lys Lys
195 200 205 Leu Ala Glu Arg Arg Pro Asp Val Pro Ile Tyr Val Gly Asn
Thr Glu 210 215 220 Arg Pro Val Phe Trp Asn Leu Asn Gln Ser Gly Val
Gln Leu Thr Asn 225 230 235 240 Ile Asn Val Val Pro Phe Gly Ile Trp
Gln Gln Val Asp Lys Asn Leu 245 250 255 Arg Phe Met Ile Leu Met Asp
Gly Val His Pro Glu Met Asp Thr Cys 260 265 270 Ile Ile Val Glu Tyr
Lys Gly His Lys Ile Leu Asn Thr Val Asp Cys 275 280 285 Thr Arg Pro
Asn Gly Gly Arg Leu Pro Met Lys Val Ala Leu Met Met 290 295 300 Ser
Asp Phe Ala Gly Gly Ala Ser Gly Phe Pro Met Thr Phe Ser Gly 305 310
315 320 Gly Lys Phe Thr Gly Asn Ser Leu Tyr Gln Asn Asp Ala Lys Glu
Leu 325 330 335 Ala Trp His Phe Ala Lys Cys Cys Val Asn Gln Tyr Lys
Asp Asn 340 345 350 9 601 DNA Porcine 9 ctgccagcct aagccacagc
cacagcaacg ctgggtctga gccatgtctg cagcctatgc 60 cagagctccc
cgcagcgccg gatgcttaac ccactgagca aggccaggga ttgaaccctc 120
gtcctcatgg atagcagttg agttgtttcc acggaactct taggggaact cctgattatt
180 ttttatttaa atttatattt ctctgacttt ttcgtgtgct catcagccac
tgactgtgta 240 tctccattag tcatggtttg ttaactctgt cattcaaacc
ctcttcatcc ttgctacgca 300 gataacatca ttataataaa atcgtgcctg
aagaccagtg acgcccccaa gctaagttac 360 tgcttcccct ggggggaaaa
agaagcaccg cgcgggcgct gacacgaagt ccgggcagag 420 gaagacgggg
cagaggaaga cgggggagca gtgggagcag cgggcagggc gcgggaagca 480
ctggggatgt tccgcgttgg caggagggtg ttgggcgagc tcccggtgat gcagggggga
540 ggagcctttt ccgaagtagc gggacaagag ccacgggaag gaactgttct
gagttcccag 600 t 601 10 228 DNA Porcine 10 cccgacgtcc tggcagcgcc
caggcactgt tattggtgcc tcctgtgtcc acgcgcttcc 60 cggccaggca
gccctggcgg atcctatttt ctgttccccc gattctggta cctctccctc 120
ccgccctcgg tgcgcagccg tcctcctgca gtgcctgctc ctccaggggc gaaaccgatc
180 agggatcagg ccacccgcct cctgaacatc cctccttagt tcccacag 228 11 98
DNA Porcine 11 tctaatgcct tgtggaagca aatgagccac agaagctgaa
ggaaaaacca ccattctttc 60 ttaatacctg gagagaggca acgacagact atgagcag
98 12 204 DNA Porcine 12 catcgaacaa acgacggaga tcctgttgtg
cctctcacct gccgaagctg ccaatctcaa 60 ggaaggaatc aattttgttc
gaaataagag cactggcaag gattacatct tatttaagaa 120 taagagccgc
ctgaaggcat gtaagaacat gtgcaagcac caaggaggcc tcttcattaa 180
agacattgag gatctaaatg gaag 204 13 92 DNA Porcine 13 gtctgttaaa
tgcacaaaac acaactggaa gttagatgta agcagcatga agtatatcaa 60
tcctcctgga agcttctgtc aagacgaact gg 92 14 110 DNA Porcine 14
ttgtagaaaa ggatgaagaa aatggagttt tgcttctaga actaaatcct cctaacccgt
60 gggattcaga acccagatct cctgaagatt tggcatttgg ggaagtgcag 110 15
191 DNA Porcine 15 atcacgtacc ttactcacgc ctgcatggac ctcaagctgg
gggacaagag aatggtgttc 60 gacccttggt taatcggtcc tgcttttgcg
cgaggatggt ggttactaca cgagcctcca 120 tctgattggc tggagaggct
gagccgcgca gacttaattt acatcagtca catgcactca 180 gaccacctga g 191 16
148 DNA Porcine 16 ttacccaaca ctgaagaagc ttgctgagag aagaccagat
gttcccattt atgttggcaa 60 cacggaaaga cctgtatttt ggaatctgaa
tcagagtggc gtccagttga ctaatatcaa 120 tgtagtgcca tttggaatat ggcagcag
148 17 82 DNA Porcine 17 gtagacaaaa atcttcgatt catgatcttg
atggatggcg ttcatcctga gatggacact 60 tgcattattg tggaatacaa ag 82 18
138 DNA Porcine 18 gtcataaaat actcaataca gtggattgca ccagacccaa
tggaggaagg ctgcctatga 60 aggttgcatt aatgatgagt gattttgctg
gaggagcttc aggctttcca atgactttca 120 gtggtggaaa atttactg 138 19
1014 DNA Porcine 19 gtaattcttt atatcaaaat gatgccaagg agttggcatg
gcactttgct aaatgctgtg 60 tgaatcaata caaagataat taggacatgg
ttcttcctca caagaggtgt gcaatcttat 120 tgggaaatca tacttgcaag
tcacaaatat agactaaagt ttccagctga gaatatgctg 180 atggagcatg
aaacactaag gagacaggga gaatctcagg aaaaatcaag aataatttgg 240
atcaaatgga ttcctgacat agaacataga gctgatcaga aagagtctga cattggtaat
300 ccaggcttaa gtgctctttg tatgtggttc agaacagagt gtgggcagcc
tgagggggat 360 acataccctt gacctcgtgg aaagctcata cgggggaggg
atgaggctaa ggaagcccct 420 ctaaagtgtg ggattacgag aggttggggg
ggtggtaggg aaaatagtgg tcaaagagta 480 taaacttcca gttacaagat
gaataaattc taggggtata ataacagcat ggcactatag 540 atagcatatt
gtactatata ctggaagtgc tgagagtaga tcttacatgt tctaaccaca 600
cacacacaca cacacacaca cacaccacac acacacacca cacacacaca cgtgcacaca
660 aacagaaatg gtaattatgt gaggtgatgg cggtgttaac taactttatt
gtggtcatca 720 tttagccata catgcatgtc atgaaatcac catgttgtac
accttaaagt tatgtaatac 780 tagatgtcag ttatatctca aagctagaaa
aaatgtgggg accaaggcag aagctcttct 840 gctctgtgtc taagggtggt
tctggggctg ggatggggag gatggttaag tggtatattt 900 ttttcatacc
tttgctcagt actatcattg taagtgttca atatatgtct gcttaataaa 960
ttaatgtttt tagtaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 1014 20
136 DNA Porcine 20 aggaatggaa agcccaattc attaaaacag aaaggaagaa
actcctgaac tacaaggctc 60 ggctggtgaa ggacctacaa cccagaattt
actgcccctt tcctgggtat ttcgtggaat 120 cccacccagc agacaa 136 21 169
DNA Porcine 21 gtatggctgg atattttata taacgtgttt acgcataagt
taatatatgc tgaatgagtg 60 atttagctgt gaaacaacat gaaatgagaa
agaatgatta gtaggggtct ggagcttatt 120 ttaacaagca gcctgaaaac
agagagtatg aataaaaaaa attaaatac 169 22 141 DNA Porcine 22
gtatattaag gaaacaaaca tcaaaaatga cccaaatgaa ctcaacaatc ttatcaagaa
60 gaattctgag gtggtaacct ggaccccaag acctggagcc actcttgatc
tgggtaggat 120 gctaaaggac ccaacagaca g 141 23 175 DNA Porcine 23
caagggcatc gtagagcctc cagaagggac taagatttac aaggattcct gggattttgg
60 cccatatttg aatatcttga atgctgctat aggagatgaa atatttcgtc
actcatcctg 120 gataaaagaa tacttcactt gggctggatt taaggattat
aacctggtgg tcagg 175 24 117 DNA Porcine 24 atgattgaga cagatgagga
cttcagccct ttgcctggag gatatgacta tttggttgac 60 tttctggatt
tatcctttcc aaaagaaaga ccaagccggg aacatccata tgaggaa 117 25 31 DNA
Porcine 25 atcctgtgtc aggagttggg attctttgaa g 31 26 116 DNA Porcine
26 attcggagcc gggttgatgt catcagacac gtggtaaaga atggtctgct
ctgggatgac 60 ttgtacatag gattccaaac ccggcttcag cgggatcctg
atatatacca tcatct 116 27 97 DNA Porcine 27 gttttggaat cattttcaaa
taaaactccc cctcacacca cctgactgga agtccttcct 60 gatgtgctct
gggtagagag gacctgagct gtcccag 97 28 655 DNA Porcine 28 gggtgcccaa
caacatgaaa aaatcaagaa tttattgctg ctacgtcaaa gcttatacca 60
gagattatgc cttatagaca ttagcaatgg ataattatat gttgcacttg tgaaatgtgc
120 acatatcctg tttatgaatc accacatagc cagattatca atattttact
tatttcgtaa 180 aaaatccaca attttccata acagaatcaa cgtgtgcaat
aggaacaaga ttgctatgga 240 aaacgagggt aacaggagga gatattaatc
caagcataga agaaatagac aaatgagggg 300 ccataagggg aatataggga
agagaaaaaa attaagatgg aattttaaaa ggagaatgta 360 aaaaatagat
atttgttcct taataggttg attcctcaaa tagagcccat gaatataatc 420
aaataggaag ggttcatgac tgttttcaat ttttcaaaaa gctttgttga aatcatagac
480 ttgcaaaaca aggctgtaga ggccacccta aaatggaaaa tttcactggg
actgaaatta 540 ttttgattca atgacaaaat ttgttattta ctgcggatta
taaactctaa caaatagcga 600 tctctttgct tcataaaaac ataaacacta
gctagtaata aaatgagttc tgcag 655 29 363 DNA Porcine 29 gtgagaaggc
ttcgccgctg ctgccgctgg cgccggcagc gccctccacg cacttcgtag 60
tgggcgcgcg ccctcctgca ttgtttctaa aagatttttt tttatccgct tatgctatca
120 gttactgagg aagtatttac aaatctacta ttattttgaa tttgcctttt
tctccttata 180 gtttatcagt atctcttgag actgttattg gtgcctgcaa
atttaaaatg attggggttt 240 tatgaggaag tgaacctttt atctttatga
aacgcctaac tgaggcaatg ttaattgctt 300 aaaatacttt ctttattatc
agtgtggcca tgccagtgtc ctcttggtta gaatttgcct 360 gat 363 30 143 DNA
Porcine 30 ctgccaaagc tgggagatgg gggaaagtag agtgggttat tgaaactgaa
tatagagttc 60 agcatctaaa agcgaggtag tagaggagga agctgtgtca
acggaaatac tgagctgggt 120 tcacatcctc tttctccaca cag 143 31 237 DNA
Porcine 31 gcaagtgaga gggggcttta gctgtcaggg aaggcggaga taaacccttg
atgggtagga 60 tggccattga aaggagggga gaaatttgcc ccagcaggta
gccaccaagc ttggggactt 120 ggagggaggg ctttcaaacg tattttcata
aaaaagacct gtggagctgt caatgctcag 180 ggattctctc ttaaaatcta
acagtattaa tctgctaaaa catttgcctt ttcatag 237 32 4229 DNA Porcine 32
gtactgagaa tcctttgctt tctccctggc gatcctttct cccaattagg tttggcagga
60 aatgtgctca ttgagaaatt ttaaatgatc caatcaacat gctatttccc
ccagcacatg 120 cctaactttt tcttaagctc ctttacggca gctctctgat
tttgatttat gaccttgact 180 taatttccca tcctctctga agaactattg
tttaaaatgt attcctagtt gataaacagt 240 gaaacttcta aggcacatgt
gtgtgtgtgt gtgtgtgtgt gtgtgtttac cagcttttat 300 attcaaagac
tcaagcctct tttggatttc ctttcctgct ctctcagaag tgtgtgtgtg 360
aggtgagtgc ttgtccaaac actgccctag aacagagaga ctttccctga tgaaaacccg
420 aaaaatggca gagctctagc tgcacctggc ctcaacagcg gctcttctga
tcatttcttg 480 gaagaacgag tgctggtacc ccttttcccc agccccttga
ttaaacctgc atatcgcttg 540 cctccccatc tcaggagcaa ttctaggagg
gagggtgggc tttcttttca ggattgacaa 600 agctacccag cttgcaaacc
agggggatct gggggggggg tttgcacctg atgctccccc 660 actgataatg
aatgagggat tgaccccatc ttttcaagct ttgcttcagc ctaacttgac 720
tctcgtagtg tttcagccgt ttccatatta ggcttgtctt ccaccgtgtc gtgtcgtcaa
780 tcttatttct caggtcatct gtgggcagtt tagtgcgaat ggactcagag
gtaactggta 840 gctgtccaag agctccctgc tctaactgta tagaagatca
ccacccaagt ctggaatctt 900 cttacactgg cccacagact tgcatcactg
catacttagc ttcagggccc agctcccagg 960 ttaagtgctg tcatacctgt
agcttgcttg gctctgcaga tagggttgct agattaggca 1020 aatagagggt
gcccagtcaa atttgcattt cagataaaca acgaatatat ttttagttag 1080
atatgtttca ggcactgcat gggacatact tttggtaggc agcctactct ggaagaacct
1140 cttggttgtt tgctgacaga ctgcttttga gtcccttgca tcttctgggt
ggtttcaagt 1200 tagggagacc tcagccatag gttgttctgt caccaagaag
cttctgcaag cacgtgcagg 1260 ccttgaggtc ttccgacttg tggcccgggg
actctgcttt ttctctgtcc ttttttctcc 1320 ttagtgggcc atgtcctgtg
gtgttgtctt agccagttgt ttaagggagt gttgcagctt 1380 tatgattaag
agcatggtct ttccttgcaa actgcttggt ttagaagcct ggctccacca 1440
cttagcggct ctgtgacctc ggacacattt cttagccttt ctgggcctcg ctcttcttcc
1500 tcataaagtg aaaatgaaag tagacaaagc cttctctgtc tggctactga
gaggatggag 1560 tgatttcata cacataaagc acttaaaata atgtctggca
tatgatacat gctcaataaa 1620 tgtcacttac atttgctatt attattactc
tgccatgatc ttgtgtagct taagaacaga 1680 ggtctttaca ggaattcagg
ctgttcttga atctggcttg ctcagcttaa tatggtaatt 1740 gctttgccac
agactggtct tcctctcctt cacccaaagc cttagggggt gaacgatccc 1800
agtttcaacc tattctgttg gcaggctaac atggagatgg caccatctta gctctgctgc
1860 aggtggggag ccagattcac ccagctttgc tcccagatac agctccccaa
gcatttatat 1920 gctgaaactc catcccaaga gcagtctaca tggtacactc
ccccatccat ctctccaaat 1980 ttggctgctt ctacttaggc tctctgtgca
gcaattcacc tgaaatatct cttccacgat 2040 acagtcaagg gcagtgacct
acctgttcca ccttcccttc ctcagccatt tttcttcttt 2100 gtacataatc
aagatcagga actctcataa gctgtggtcc tcattttgtc aatctaattt 2160
cacagcctct tggcacatga agctgtcctc tctctccttt ctgcctactg cccatgagca
2220 gttgtgacac tgccacattt ctcctttaac gacccagcct gctgaatagc
tgcatttgga 2280 atgttttcaa tttttgttaa tttatttatt tcatcttttt
tttttttttt tttttttttt 2340 ttttttaggg ccgcacccat gggatatgga
ggttcccagg ctagggatcc aatgggagct 2400 gtagctgctg gcctacacca
cagccacagc aatgcacaat tcgagccaat ctttgaccta 2460 caccagagct
cacggcaaca ctggattctt aacccactga ttgaggccag ggatcaaact 2520
ctcgtcctca tagatacgag tcagattcgt taacctctga gccatgatag ttgttagtta
2580 ctcattgatg agaaaggaag tgtcacaaaa tatcctccat aagtcgaagt
ttgaatatgt 2640 tttctgcctt gttactagaa aagagcatta aaaattcttg
attggaatga agcttggaaa 2700 aaatcagcat agtttactga tatataagtg
aaaatagacc ttgttagttt aaaccatctg 2760 atatttctgg tggaagacat
atttgtctgt aaaaaaaaaa aatcttgaac ctgtttaaaa 2820 aaaaaacttg
actggaaaca ctaccaaaat atgggagttc ctactgggac acagcagaaa 2880
tgaatctaac tagtatccat gaggacacag gtttgatgcc tggcctcgct aagtgggtta
2940 aggatatggt gttgctgcag ctccaattca acccctatcc tgggaacccc
catatgccac 3000 cctaaaaagc aaaaagaaag gtgctgccct aaaaagcaaa
aagaaagaaa gaaagacagc 3060 cagacagact accaaatatg gagaggaaat
ggaactttta ggccctatct ccaactatca 3120 catccctatc accgtctggt
aagaaatgga aaaaatatta ctaagcctcc tttgttgcta 3180 caattaatct
gattctcatt ctgaagcagt gttgccagag ttaacaaata
aaaatgcaaa 3240 gctgggtagt taaatttgaa ttacagataa acaaattttc
agtatatgtt caatatcgtg 3300 taagacgttt taaaataatt ttttatttat
ctgaaattta tatttttcct gtattttatc 3360 tggcaaccat gatcagaaat
ctttaaacaa tcaggaagtc ttttttctta gacaaatgaa 3420 aatttgagtt
gatcttaggt ttagtacact atactagggg ccaagggtta tagtgtgact 3480
attaaatcac agataatctt tattactaca ttatttcctt atactggccc cacttggatc
3540 ttacccagct tagcttttgt atgagagtca tccttaaaga tgactttatt
ctttaaaaaa 3600 aaaaacaaat tttaagggct gcacccatag catatagaag
ttcctaggct agcggtcaaa 3660 ttagagctgc agctgccagc ctatgccaca
gccacagcaa tgccagatct gagctgcatc 3720 tgtgacctac actgcagctt
gcagcaatgc tggatcctta acccattgaa caatgccagg 3780 gattgaacac
acatcctcat ggatactgct caggttccta acctgctgag ccacagttgg 3840
aactccaaag cagactttat tctgatggct ctgctgatct ctaacacgtt attttgtgcc
3900 atggtgttta tcttcacttt actcaagtca gggaaacacg aagagtctca
tacaggataa 3960 acccaaggag aaatgtgcaa agtcacatac aaatcaaact
gacaaaaatc aaatacaagg 4020 aaaaaatatc ttcactttca aaatcaccta
ctgatgatga gtttatattt ccttggatat 4080 ttgaatatta gctatttttt
tcctttcatg agttttgtgt tcaaccaact acagtcgttt 4140 actttgatca
cagaataatg catttaagcc ttaaatagat taatatttat tttcaccatt 4200
tcataaacct aagtacaatt tccatccag 4229 33 8164 DNA Porcine 33
gtaaatacca tcaatactga tcaatgtttt ctgctgttac tgtcattggg gtccctcttg
60 tcaacttgtt tccaatctca ttagaagcct tggatgcatt ctgattttaa
actgaggtat 120 tttaaaagta accatcactg aaaattctag gcaagttttc
tctaaaaaat cccttcattc 180 attcatttgt tcagtaagta tttgatgaga
ccttaccatg tgtaaacatt gcactaggta 240 ttaagaaata caaagatgga
taagatagag tcggcgtaaa tgagatgata taatgagacg 300 ttataatgaa
actcacaatt ccagttggga aataaagtcc ttcaaattcc atgactcttt 360
ctggcacacg ttagaggcta cagcttctgt gtgattctca tgctggctcc acttccactt
420 tttccttctt cctactcaag aaagcctata gaaatatgag taagaagggc
ttaatcatag 480 gaataaattt gtctctgttc taagtgatta aaaatgtctt
tatcagtata aaaagttact 540 tgggaagatt cttaaaactg cttttacaca
ctgttctaga atgactgtta tataaataaa 600 aaagtagatt tgatctaaca
caattaaatg acctttggaa atattgacta attctcacct 660 tgcccctcaa
agggatgcct gaaccatttc cttcttttgc cagaaagccc ccaccctttg 720
tctgttgacc tagcctagga aatcttcaga tcacgttgtt agcacgaact ggttacatgt
780 gctgtacaaa tactatttaa ttcatctgat taaaaaaaaa gagataagaa
gcaaaagttt 840 gactatctta aactgtttgc gtaggtgaga ggacaattga
ccatctactt tatgagtatg 900 taacccagaa acttaaagct ccttaaggga
gctaagtctt ttggataaga cctatagtga 960 gaccttttag caaaatggtt
aagactgaat ggagctcact agcgtgggtt catatcctga 1020 tgctcaaaca
cgcaattaaa tgactttagg tgggttagtc tctgttcctt agtttcctca 1080
atgggagata atattggtag tagcgatttt actgggttgt tgaaagaaca tctgttaaat
1140 gttcagaacg tgttacgaca gagtacagag taatgatttg cttgtatatg
tatgactcaa 1200 atagtctgcc atatgccttg tgactgggtc ctgtggagca
ggaaggaggg atttcccacc 1260 cagcagaaag ttgggtaaac tggaaaatag
actgaggcca ggaaatgatg caaagcgttg 1320 atgttcactg ccacggcagg
tgaagggcag ggccagagtt gtcagtaggg tcaggggagg 1380 actggaaata
accaagaccc actgcacttt tcagcctttg ctccagtaag gtaatgttgt 1440
gagagtagaa aattttgtta acagaaccca cttttcagta cagtgctacc aatactgtag
1500 tgatttcata ccacatccca agaaagaaaa agatggctca atcccatgtg
agctgagatt 1560 atttggtttt attgttaaat aaatagcatt gtgtggtcat
cattaaaaaa ggtagatgtt 1620 aggaaagtag aaggaagaag actctcacct
acattttcat cactgttttg gtatctgcca 1680 gttgtcacct tggtcccctt
ccccgcctct cccctgcctc ctcttcctcc ttctcctttt 1740 tttggaatac
aattcaggta ccataaaatt taccctttta gagtgtttga ctcaatggtt 1800
tttagtattt tcacatgttg tgctattact atcactatat aattccaggt cattcacatc
1860 accccccaaa gaaaccttct aactattagc agtccattcc cttcttccct
cagcccctgg 1920 caaccactaa tctacttact gtctccatgg atgttcctat
attgaatcaa gctagcataa 1980 accccacttg ctcatggtca taattctttt
ttatagtgct aaattacatt tgctaatatt 2040 caattaagga tttctatgtc
catattcata aggaatattg gtgtgtagtt ttctctttgt 2100 gtgatatctt
tgtctggttg ggggatcaga gtaataatta ctgctctcat agaatgaatt 2160
gagaagtgtt ccctcctttt ctatttattg gaagagtttg tgaagtatat tggtattgat
2220 tcttctttaa acatttggtc agattcacca gtgaagccat ctgggccatg
gctaatcttt 2280 gtgaaaagtt ttttgattac taattaaatc tctttaattt
gttatgggtc tgctcctcag 2340 acgttctagt tcttcttgag tcagttttgt
tcatttgttt cttcctagga ctttctccct 2400 ttcatttgga ttatttagat
tgatagtaat atcccccttt taattcctgg ctgtagtaat 2460 ttgggtcttt
tctctttttt cttggtcagt ttagctaaag gtttgtaatt gtattaatct 2520
tttcaaataa ctaacttttt tgttttgttt gttttttgtt ttttgttttt tgttttttgt
2580 ttttttttgc tttttaaggc tgcacctgag gcatatggaa gttctcaggc
tagaggtcta 2640 atcggagcta cagctgctgg cctataccac aaccatagca
atgccagatt caagctgcat 2700 ctgcgaccta caccacaact cggccaggga
tcacacccgc aacctcatgg ttcctagtcg 2760 gatttgttaa ccactgtgcc
acgacgggaa ctcccgccca ttttttttaa cacctcatac 2820 tttaacataa
agatgggctt cacatggact gatagctcaa atgaggaagg taagactatg 2880
aaagtaatgg aagaaatgta gactattttt gtgacctaga gattactgat acttcttgac
2940 ttttcaaaca atacttcaaa agtacagccc aaagggaaaa aagaaagaaa
aaagaaacac 3000 acatatacac aaacctagtg aataagatat catcgataca
ctacagattt ctatgaactg 3060 gaagacccca tggacaaagt taaagaacat
atgatagttt gagtgattat tttgcaatat 3120 ttacaaccaa tgagggaata
ttatccagct tataggagga agtaatgcaa atcgacaaga 3180 aaaagatagg
aaacccaata taaaaattaa gaaaatacaa aaattaagaa aggatatgaa 3240
ctagcatttt acaaaagaaa aatctccaaa agtcaatcag cacatgaaaa tatgctcaaa
3300 cctattaatt attagaaaac tacagactga agcaatgagg tgctttactt
tacatctttt 3360 tgactgataa aaagttagaa acaaaggtga tatcaaatgt
cagggataaa aggatataga 3420 aatcgtcatg cctgtggtgg gagtatggcc
ggtgcagtca tgtgggaagg taatctgaca 3480 gtggttaggc agagcaggtt
tatgaataca ctgtggccca tcaatcccac gcctgtttat 3540 gtaccaaaga
aatcctgttg tggcagaatc tatgggtcca cccctgggag catgaattaa 3600
taaaatgtgg caccagggtg tgtgaaactc cagctagaga tgagatgtcc acatggcaac
3660 atgaatgcat cttagaaaca tagatttgag tgaaaaagag taagaaacag
ccgggaaacc 3720 caataccatt tataaaaatt aaagatgcac acatacaatg
tagtaaatat tttgcatgaa 3780 ctttcaaatg gttgcctaca gggggggaga
gtaaagaaga gtagaaaaca aagataaagg 3840 gagtaagtaa gtagctctgc
ctggactgaa tataatgtgt catgaactga gaaatatggt 3900 taacataatc
ctcttaactt gaggtcctaa atgaatgaat gagtccacta ttcatttacc 3960
cattctttaa tgtgtattgc attataatcc atttttttag aaccaacgaa ttttgttccc
4020 ataactacta atcagcctgc cttttctccc tcattccctt atcagctcag
gggcattcct 4080 agtttttcaa acgttcctca tttgaaccaa aaatagcatc
attgtttaaa ttatacttgt 4140 tttcaaatac gatgcttata tattccaagt
gtgtttgccc attttcttag gtggtagaaa 4200 tttttcattc tacttttcta
tctactcaga ttttcccgtt ggaattattt ccattgctat 4260 taaacttaga
agtcccccct gtgatatgcc atttttttca tactttttaa gcacttggtt 4320
gcttttcttt gtgtctttaa gcacctagaa tacttataac cattgcacag cactgtgtat
4380 caggcagccc ttcctcttcc actaatttat ggtccttctc ttagactata
ttaaactgtt 4440 atttaattag gatcctctct tcgtccttat gatttaatta
ttatagtttt ctaatatgtt 4500 tttattataa ttcctcttca ttattcctcc
ctattaaaaa ttttaatgaa ttccatttgt 4560 ttgttcttct agttaaatat
taagtcataa tccaaataac ttagatgtca ttagtttatg 4620 tggtcaaagt
aaggatacca catctttata gatgcaggca gttggcagat gtcatgattt 4680
tcttcagtgc ataaatgcaa tttatctttg agcaaggggc ataaaaactt ttatggtatt
4740 ggctttgaaa taatagttaa gaactgcaga ctcagttttt cctgcttttc
ttgaaaaaga 4800 acacttctaa agaaggaaaa tccttaagca tggatatcga
tgtaattttc tgaaagtctc 4860 ctgtaattcc ttgggatttt tgttgttgtt
tgttggtcgg tttttttggg tttttgtttg 4920 tttgttttgt tttgttttgt
tttgctttta gggctgcacc tgtggcatat ggaagttccc 4980 aggctagggg
tccaactgga gctacagctg ccagcctact ccacagccac agcaacatgg 5040
gatcctagct gcatctgtga cctaaccaca gctcttggta atgccagatt gttaacccac
5100 tgagcaatgc cagagatcga atctgcctcc tcatggacac tagtcagatt
agtttctgct 5160 gagccacaat gggaattccc aattccttgt atttttgaac
tggttatgtg ctagcatata 5220 attttgtttc ttgaatcttt gtgggttttt
tttttttttt ttttttgtct cttgtctttt 5280 taaggctgca cccacagcat
atggaggttc ccaggctaga ggtcaaattg gagctacagc 5340 tgccagccta
cacaacaact gcagcaaagt ggggcccaac ttatatgaca gttcgtggca 5400
atgccggatt cctaacccac tgagcagggc cagggatcga acctgagttt ccagtcagtt
5460 tcgttaacca ctgagccatg atagtaactc ctgtttgttc agtcttgaac
ctccttttta 5520 attctttatt ccttgagggt gaaataattg ccataataat
actatcattt attacatgcc 5580 ttctctgtgc taggcatagt gacactttag
gatttattat atcacttaat ccctacaaca 5640 actctgcaaa gtatgtatca
taatcctatt tgacagatca ggaaattgca gcccaggatg 5700 cagataatat
gcatccatca caagtgacta gatatagtcc ctctgctatt cagcagggtc 5760
tcattgcctt tccattccaa atgcaatagt ttgcatctat tgtatatgtg ttttggggtt
5820 tttttgtctt tttttttttt tttgtctttt ctggggcctc acccttggca
taggtaggtt 5880 cccaggctag gggtcaaatt gaagctgcag ctgccagcct
acaccacagc cacagcaact 5940 cgggatctga gcctcatctg caacctacac
caaagctcac ggcaacaccg gatccttaac 6000 ccactgagtg aggccagaga
tcaaaccggc aacctcatgg ttcctagtcg gattcattaa 6060 ccactgagcc
acgatgggaa ctccctaaat gcaatagttt gctctattaa ccccaaactc 6120
ccagtccatc ccactccctc ctcctccctc ttggcaacca caagtctgtt ctccatgtcc
6180 atgattttct tttctgggga aagtttcatt tgtgccattt ttcattttac
gggtaatttt 6240 tacttcagtt tcttccacta gcagttgtct taaagtgagt
ataattaata ttcatttgga 6300 aaatgtaagc aaaacatttt ttaaagggcc
atgcccacag catatgaaag tttctgggcc 6360 aggggttgaa tccaggctcc
aagttgcagc tgtgccctac actgcagctg ggcaatgctg 6420 gatcctttaa
cccactgtgc ccggctaggg atcaaacctg catttccaca gctacccgag 6480
ccattgcagt tggattctta acccactgca ctacagtggg aactcccaca aaacattttt
6540 taatgtcctt tgaataaagt aggaaagtgc tcgtctttga gggcagggcg
gcaatgccat 6600 ttccacaagg tttgctttgg cttgggacct catctgctgt
catttagtaa tgaataaaat 6660 tgctgacagt aataggatta actgtgtgtg
gagatagcca gggttagaga taaaaacact 6720 ggagaagtca aataagttgc
tcgaggtcct ctagctaata agctattaag tgggagagtg 6780 agggctagaa
acaggccatc tgtctcccaa gcacatgtcc attagtggtt tgctgatagc 6840
cttccagaac aacagagagg actctcaaac atggtcttgc ctccctccaa ttgatcccct
6900 ccatgtgcct cacagcgggt ctttctaaaa ttaagttctg attttaattc
tcccttgcta 6960 tagcacttag gtatggcttt cagccgtgca ataaaaagca
ggcaagagtg gctcaatcat 7020 ataggaggtt gtttttctta gatcccaagc
aggtaatcct gggcattatg gttgttctgc 7080 gtttatcaag gagccaaatt
ctctatcacc tcctgttcta tcctcctcag tatctggctc 7140 tattcttcag
catctcaaga tggcttgtgc tcctccaagc atggcagtca aattccacac 7200
aagaggggga aatatgaagg gcagacagtg ctggtctcct gagctgtccc tctttgtcgg
7260 ggaaataaat gtattccttc aagtcccgtg agacttctga agtagacgtc
tgcttacgtc 7320 tcacccacca gaactatgta aactgcacat agtgctaggt
ctacatagcc actcataact 7380 gccagggggt gggaaatctt taaataggtg
taccaccaca caattaggat gctaatagta 7440 agggagaagg agagaatagg
ttttgcgcaa gccaccagca tgcctgccac aattgcttaa 7500 aattcttcat
tgacccctca ttgccacagg atgaaatcca aacgccttct tagttgggaa 7560
tctgacctac ctgtctctcc cacctggttc agacaccatt ctccttggtc ataaaattcc
7620 agtcatttgt gaacatccag ctcccccatg cctccatgcc tttgcacatg
ctgttctttt 7680 atcttttatg ttgtcctttt atcttttatc caaaagagat
atcccatcat cacatctctt 7740 ttgtcagccc ccaaatactt tgtctttcaa
gttcagctgg aggattacct cctatttgaa 7800 atcagctttg tctcttacaa
ccaaacaagg ttttccttcc gagacactcc cacagcacct 7860 tgaactcatc
tctatcaatc attcatttga ttgtaatgaa gttgttggtg gtatgcctgt 7920
gtctctgaca catctgcgat ctcatgagtt ccttaagtgg aatgtgaata gcgggatgaa
7980 cagtattggt cttcagccct catctctgca gatgttgctt gacccaaatg
agcgttgcct 8040 tttattttga ttttgctttg atttgtctac tccatgtact
tgagccatgc atttctgtct 8100 tagcgatgct ttttaaaagt cattttttgg
ttgattatcc agatttgtcc acctttgctt 8160 ctag 8164 34 1604 DNA Porcine
34 gtaaggaaat gttaaattgc aatattctta aaaacacaaa taaagctaac
atatcaattt 60 atatatatat atatatatat attttttttt ttttttacat
cttatattac cttgagtatt 120 cttggaagtg gctagttagg acatataata
aagttattct gaagtctttt tttttctttt 180 tccatggtga gcagtggctt
gatgtggatc tcagctccca gacgaggcac tgaacctgag 240 ccgcagtggt
gaaagcacca agttctagcc actagaccac cagggaactc cctattctaa 300
attcttgagc acattattta ggaacctcag gaacttggca ggattacagg aaatatatct
360 agatttaaaa aaaaatcttt taacagaggt cccaaaggag agtcatgcac
agctatggga 420 ggaagttcag aaactgccct tgctaccaga tcactgtcag
ataaaatggc cagctacatg 480 tttctgcaca ttgccctaag atctttacaa
acttttctgt gcatttttcc acttttaaaa 540 gaaaatttcg gggttcctgt
tgttgctcag tggttaacga acccaactag tatccatggg 600 gacaggggtt
cgagccctgg cctcactcag tgggttaaga atctggcatt gctgtggctg 660
tggcgtaggc tggcggctac agctcagatt ggacccctag cctgagaacc tccatatgcc
720 gcaggtatgg ccctaaaaaa aaaaaaaaag agagagagag aatttcctcc
agaaaaaaca 780 ctttggtagt ttgggagaag taaacaacca aaaattaatt
tttctggagt attcgggaag 840 cttgtaaaaa tgggctctta cttttttgag
gagacaaatg ggaacctacc cagaagaggc 900 acaatcacct gcatttgatt
tcttgacctc tccctacctt ctttgctggc tttccacatt 960 tggatttctg
tgaccttatc tctgctcctt ggtgttttca tttttcctgt ggacgtgcca 1020
gactatggga agggagtaag gcgttgattt agaatcctgt agtctctgcc tgtctctagt
1080 cattgttttc acccttctca aaggaccttg acatcctgag tgagtccgca
agtaatttag 1140 gggagaagcc ttagaagcca gtgcagccag gctacatgac
tgtgtccacc cactggaacc 1200 agtcattttt atacctattc acagcccccc
taccatttaa atccccagag gtctgccata 1260 acatctgtaa ctccctttcc
tggtaaattg tgttctaaaa gactggtaac aaaagatatt 1320 ctgtggtaca
gagcataatt aaatacctgg gagctgattt gagtggggta aatcaactgg 1380
tttgacccct aaaacccacc atgagcattt ctgttctaat aaagtaatgc ccgtgctggg
1440 aattgtgttc tacggaaatg ctcctgctgt gtctttcttg agtcctgtgt
cattgaacat 1500 gcttaggagc aaaggtcccc catgtggctt gtctgctaac
cagcccagtt ccttgttctg 1560 gctggtaatg atccgatcat ctgaatctca
ctgtcttcca acag 1604 35 3628 DNA Porcine misc_feature (361)..(361)
n is a, c, g, or t 35 gtaaggaagg gtgagccctc aactccgaag aaaatgctgc
aataaaagca ctgttggttt 60 tcagcttttt ttgtaatcac tgctcattct
gaggtagatt cgcttgggct gataaaaaga 120 gaactaattc agataaatgc
ttgcatttgc atagcctctt tttttaaaaa cttttttttt 180 tttttttttt
ttttggcttt tcagggctga acctgtggca tatggaggtt cccaggctag 240
gggtcgaatc agagctgtag ccccgggcct atgccactgc catagcaaca tgcatagcct
300 cctttttaaa gtgccttcct gttttatacc attgggatgt gagaagagct
attgtggaaa 360 ngagcatggg gtnataaccc tggacctctc acgtcctacc
ctcaggntag tgggaaaact 420 ctgagtttaa ggacatcaaa gtgactcctt
tttagttaca ttatggngga atcagcncat 480 atttttacaa ggggcggagn
gtaanctgtt ggagtttaca agacatatgg tggcattgca 540 actacttaac
cctactatta tagcacaaaa gcagccatag tcggtcctga aggagcctga 600
tgccttcagc tttataggca atgacgtgtg aatatcacaa acagtttcct gtgtcaccaa
660 acatgattgc cttttgattt ccctttcaac cctttaaaaa aaggtaaaag
cccttcttag 720 cattcagcag caggtcgctg tgttttgcca actcctgatc
tgtagcattt cgacaacact 780 gagctctcaa cttttgaacc ctgagtccac
cacatccttc agtgaaacca gagccatgtg 840 atactaagga tagaaacgga
aacttcctga atccaggcga tcaaatagga gggagaaaga 900 ggaactttca
ttgacaaaac cacaaatatt gtgaatggac tgttacaaat attgtgaatg 960
ctcctattcc caaccccctg gcttcattac agggtcctat gtgttcatcc ttattgagaa
1020 atttgtattg ctactgccag gttgccaata cccagcggtg cccatggtgt
tctaaaatga 1080 agcaatttca actttatttt tttttcctgt gactttacat
gacaagttca catgaaggat 1140 atactttgat agtaatgtcc atggttaggg
aatatacatt gtttgctggt tgactggccc 1200 ctggattttt ctattgaaag
tccatgagat ctcgaaggca caggtgtgtt ctctcgcttt 1260 ttaaggaaag
ggtttaaaaa cttaagtaat taacagcttt agtaacaaat tacctataac 1320
acacttaaaa accgaatacc acccactgga gtattgtgct acgattaaaa atctacttgt
1380 ctactacatg atatctttgt cccacagaag gttctggaac caaacttgta
atttcaggat 1440 tatgagagcc ctgagttcac gcattgtgta ataactatgt
tgtgtggtag tcaatttgta 1500 cagcttgctt agagagaaca atgtcaagtt
aaggaggcga ttgctttata gtgcctgtca 1560 caagatgcca ttgccattgt
cctagcaaga gatattctat gggagtatac tacattttag 1620 tgaggataag
aactttttat ggcatttagt ccggtcattt cccaaccact gtcctgaaaa 1680
ccaatttcat tttgatttca ggggcttgtg tgggcaaagt tgccaggcat taaaaagcca
1740 cttctcaact gtagtatcac aatgctttag ttgggtagtg tattgcagat
agcttatggc 1800 tgaaaagtta ccaagccttg cagttttcac tcctttgagt
ttatttcctt gacagaattg 1860 accctgagtt ttttgactct tacctgctca
actaataaac accagagtca tttatctcca 1920 ttgctcttgt ctgaccttta
tttaccgaat aatgccttat gggttcacaa aaacaagggg 1980 ggagggggcc
agcatgcctt agaaactgtc tttagtcaag aaatgngatt ttattatgta 2040
aatatatgag tattataata gatagtgtta ttaatagaca ccagcaagaa ttgtcaataa
2100 tttaaaaatc acaaattaaa atacatccat gttagnatca tttatcctaa
ctcccaaagc 2160 cctttaaagt ggaagattta gatgttaacc cagagattaa
agacatgttc aaagaatcct 2220 tgattttttt ttgaatccct tgtttttaga
gaagaaaacc taatgatttt ccccctctgg 2280 attctacata ttaaatatag
ttttggaact tgaatattag tatggttaat aagtgctgat 2340 atgctgattt
tgtttatatt tttcttatga gtaaatatcc tatatcacca gacattatag 2400
tctatgtaca aatatgattc ttaaacctga tagcacattc attagagttg gaattgcctt
2460 tttttttttt ttttttacag ttgcacctgc aacatatgaa agttcccagg
ctaggggttg 2520 aatccaagct gcagctgcca ccctacatta cagccgtagt
aacagcagat ccgagctgca 2580 tctgcaacct atgctgcagc tcagggcaat
gccagatcca ctgagtgaag ccagggatgg 2640 aacttgcatc ctcatagaga
caacgtcgtg tccttaaccc actgagccag aacaggaact 2700 ccagaatttc
ctttcaatag aagaagcacc aagtttagga tcagaaagcc tgaatttgaa 2760
taccaattta ctatttgtta gtcatatatt tctgagtgtg tttcctcatt tattaaaagc
2820 agactaaaag atgagagggt cttttgttga gaatcaaata caataacatg
tgaaagtgtg 2880 taacactatg attgaaatat acctacacag ccatttattt
gtttattgtt catgttttgc 2940 cacccacaca gtagtatata atccttttat
gtaataaatg ctaataatga aagttggcaa 3000 cttatgtaag tactcaaaat
gctggaggtc atgggatact gactgggata ctacagaggt 3060 aatgtcattt
cctctgcgct aaacttattg tcttgtagtt agggactgac tctctttagg 3120
acaaggagtt cattctgtat accatgtgtg gctatcaccc ttcgaagttg aaaaactgcc
3180 ccagggtggg cacccatccg ttctcttaga tatatggccg agacctttct
ctcactggga 3240 gggaaccaca ctgaggaatg agaaaaaaaa aaggaaaatc
aagatgaaac cagaaacctc 3300 tttggcataa cttctccact ctgtactttt
tgttagaact acccttgcac aaagcagcat 3360 cagtgtggaa gacagaattt
gcacacctgg tttgatatac atgccgtggt atatgggatg 3420 ttctaacaat
aaagaggact ctcccaggaa atctcctcac tgttatagtc agccttgagg 3480
aaagagctct tcttttggac tctggggaga gtctagtttt tcagttcctt gcttctcggt
3540 caacgtgttg gtgtaaggat cacactctct cttatactag ataattctat
tttttcacct 3600 ttcaacctgt ctatccttct gaccctag 3628 36 300 DNA
Porcine 36 gtctgtgttc tttccacatg tttgggttat cctttctggg ataaatttga
ggcgagatag 60 aaactttaag actaaagaaa caatggccta ctttttttgt
acatggtcct gtgtaaatct 120 ctatttgagc tgaaataaga tggtcttcct
ctccaattat ccatggtatg actctgatgg 180 ataacaaatc cagttctgaa
aaaaggggat ttctttccag aagagaggac agtttcttca 240 aatattgaat
taaaagcaaa atagatgtaa accgttgttg gttttattgt tgaattccag 300 37 734
DNA Porcine misc_feature (705)..(705) n is a, c, g, or t 37
gtattttctt gccctcatca gcatgaaatt gctcttggta gaaaggataa
taatagttat 60 ccaaaacatc atcctatgtt catctgtttc ttccctcttc
attttccata gagtacagta 120 tattctatct ctgtcttagg aaaatggact
gtcattcata taatcttaca gagaatcaat 180 tagtaatgta ctctatgccg
tgacaggtgc gaaggttttt tttgaaggca acagataaaa 240 atatcctata
tttcacctat tgtaatttcc ttaaaactga cattattgaa taaatgtttt 300
actttcatct tgaatattat tatgttatgg aatcatacac tttaccccaa taatcatcga
360 aaagaatttc caaaaggttg agagagttgt gttgatctga ttactttcct
ctgcatcctt 420 tgagcttaac ctttgaatat agtttgctaa ggaaagtagt
ctgtttatga tcctggagtg 480 gaatcaggct aagtgtcctc attcagaacc
cactgaatca gacagaatga atttatttcc 540 ttgaaagttc aaaatgtgtc
actcaagagt ataaattttc aaatcttact ctctcttttc 600 cttggatgtg
agcaattctt cgataattga atgaggcaga ttatatagac ttacatggaa 660
gactgttggc ctgagaattc aaactatggt gttcaagact tcacngngag tccgatgcca
720 tttgtttccc acag 734 38 2421 DNA Porcine misc_feature
(1387)..(1387) n is a, c, g, or t 38 gtaattcttt atatcaaaat
gatgccaagg agttggcatg gcactttgct aaatgctgtg 60 tgaatcaata
caaagataat taggacatgg ttcttcctca caagaggtgt gcaatcttat 120
tgggaaatca tacttgcaag tcacaaatat agactaaagt ttccagctga gaatatgctg
180 atggagcatg aaacactaag gagacaggga gaatctcagg aaaaatcaag
aataatttgg 240 atcaaatgga ttcctgacat agaacataga gctgatcaga
aagagtctga cattggtaat 300 ccaggcttaa gtgctctttg tatgtggttc
agaacagagt gtgggcagcc tgagggggat 360 acataccctt gacctcgtgg
aaagctcata cgggggaggg atgaggctaa ggaagcccct 420 ctaaagtgtg
ggattacgag aggttggggg ggtggtaggg aaaatagtgg tcaaagagta 480
taaacttcca gttacaagat gaataaattc taggggtata ataacagcat ggcactatag
540 atagcatatt gtactatata ctggaagtgc tgagagtaga tcttacatgt
tctaaccaca 600 cacacacaca cacacacaca cacaccacac acacacacca
cacacacaca cgtgcacaca 660 aacagaaatg gtaattatgt gaggtgatgg
cggtgttaac taactttatt gtggtcatca 720 tttagccata catgcatgtc
atgaaatcac catgttgtac accttaaagt tatgtaatac 780 tagatgtcag
ttatatctca aagctagaaa aaatgtgggg accaaggcag aagctcttct 840
gctctgtgtc taagggtggt tctggggctg ggatggggag gatggttaag tggtatattt
900 ttttcatacc tttgctcagt actatcattg taagtgttca atatatgtct
gcttaataaa 960 ttaatgtttt tagtaagtaa tctctgttta gtaatgtgtc
agaaatgccc tacttgcaat 1020 aggaagaaaa cctgtccagt cccttccttt
tttctgtaag tctgatttca ttgcctccca 1080 gaatgcatca ccatgtgaga
gatagaggga aggtgctgtc cttatggggt taacagtgtg 1140 actagggagg
caaaatatac ctactaaagg gtggtagcat aattcagttc ttatgtgagt 1200
atgtgtatgt gtgtgagtat gtgcacatgc acatacattt taaaaggtct gtaatatact
1260 aacatgttca tagtggttac acctagctta taggtaacat tttttcccct
gtatccttgt 1320 ttgtgtttat caaattttca taacagtaat ggtagaagga
gtacctgaca tggtaccata 1380 catgctnggn cctgcctaat ttctcnattt
cctttattgc ccataccccc attgcttgac 1440 aagcataagt ccatactggc
ttgttttcgt tcctcagact cagtacacca tgtagctcca 1500 tgccctgggt
ctttgtatgt gctatttcta ctgcttagag tgctattgcc cctgaccacc 1560
acgtggtcag caacttctct tctgcgtctg tgtctatggt ctatgattcc agatgtcatc
1620 ttcactaact acccttctaa tatgcccttc catcccaccc gtcctcatcc
ttaccccagc 1680 cactctctat ttggtggctc tgttttattt tcttcctagc
tcatcactct ttgaaatgaa 1740 cttatttact tattcaatat ttgcttcttt
cactagaatg aatgctccat gagagcaggg 1800 acctgcttta tcttgctcgc
cactgtattc acagtgccta gaactacgtc tggcacatag 1860 taggtgctca
ataaatatcg atcaaatgaa agaatgagca aacgaacaaa tgaacaacac 1920
gtgaggtagg catcatgatt ccatcaacag aggagaaaac cagacttaaa gnaatgaagt
1980 ggnggagctg catttgatct tgactgactc caacatccat gctcttgacc
actgtgcatc 2040 tccagagtgt aatgaacata ctttactttt atattccacc
aaaataacaa agccatgccc 2100 atgttagtag agagttaatc gacagtgccc
ttaaaatatg catgcaccca gggtacaact 2160 atgcatgctg ccctgtgttt
tcagttggat ccaaatgaat tgccgtaaac aaagagggga 2220 ttcaatgtct
ttgactagtt tgggatattt tcctagtaac caactttgca aaataaagcc 2280
actaatgaca aggagctttg ttctacttct gcatcactca actgtcaatt tttatctctt
2340 gcaagacttc taatctacta gaacttttgt ttttctgtga tttctgaaca
gagaagacta 2400 atccaaaccc tgtcattcca g 2421 39 457 DNA Porcine 39
gtatggctgg atattttata taacgtgttt acgcataagt taatatatgc tgaatgagtg
60 atttagctgt gaaacaacat gaaatgagaa agaatgatta gtaggggtct
ggagcttatt 120 ttaacaagca gcctgaaaac agagagtatg aataaaaaaa
attaaataca agagtgtgct 180 attaccaatt atgtataata gtcttgtaca
tctaacttca attccaatca ctatatgctt 240 atactaaaaa acgaagtata
gagtcaacct tctttgacta acagctcttc cctagtcagg 300 gacattagct
caagtatagt ctttattttt cctggggtaa gaaaagaagg attgggaagt 360
aggaatgcaa agaaataaaa aataattctg tcattgttca aataagaatg tcatctgaaa
420 ataaactgcc ttacatggga atgctcttat ttgtcag 457 40 4012 DNA
Porcine 40 gtttgacttg aatatttaca gggaacaaaa atgatttctg aattttttca
tgtttatgag 60 aaaataaagg gcatacctat ggcctcttgg caggtccctg
tttgtaggaa tattaagttt 120 ttcttgacta gcatcctgag cttgtcatgc
attaagatct acacaccacc ctttaaagtg 180 ggagtcttac tgtataaaat
aaactattaa ataagtatct ttcaactctg gggtgggggg 240 ggagactgag
ttttttcaca gtcctatata ataattttct tatcctataa aataattagg 300
agttcccgta gtggctcagc aatagcaaac ccgactagta tcgatgagga tgcgggttcg
360 attcctggcc cccctcagtg ggttaaggat ctggcattgc cgtgagctgt
ggtgtaggtg 420 gcagacacgg ctcagatccc acgttactgt ggctgtggca
taggccagca gctccagctc 480 tgattagacc cttagcctgg gaacttccat
atgctgtggg tgtggccttg aaaaaaaata 540 aataaataag ataattactc
aaatgttttc cttgtctcag aaccttactt caggataaag 600 agtgagaaag
ttttttttat gaagggccat tattacagct caaaaataag ttgtcttcag 660
caagtagaaa gcaataagcc tgagagttag tgttcctatc agtgtaaata ttacctcctc
720 gccaatcccc agacagtcca tttgaacaat taacggtgcc ctgggagtac
agttcagaaa 780 cattaatgtg gatgttccag acctgtattt ttataagtac
ttgtcttgag ccggatggaa 840 ccatcattcc tcaccattat ttagaagtgg
actgtgactc tgttggagat cagggcacac 900 ggttaccaaa agcacaccct
tctcctggcc ttacctttgc aaagctgggg tctgggacac 960 agtcagctga
ttataccctt ttactaactt cccacagctc aaatctggtc aattctcctt 1020
cacaaatctc ttaaaaatcc atcactcacc tccagcctct tctgctgtgg ccttgattca
1080 gcctctcaca attttttttt aaccagaatt ctggcagtgg cccctgactt
gcctctgtgc 1140 tcccagcccc gctgtcctct gatccatcct ccatgccagc
ctttttcaat ctgctggtca 1200 cgattcattg atgggttagg aaatcaatgg
catcacaact agcatttaga aaaaggaaat 1260 aggcgttccc gccgtggcac
agcagaaata aatccgacta ggaaccataa ggttgcgggt 1320 tcaacccctg
gccttgttca gtgggttaag gatccggcat tgccgtgggc tgttttgtaa 1380
gtcacagaca tggctctgat ccggcattgc tgtggctctg gcgtaggcct gcagcatcag
1440 ctccaattag acccctatcc tgggagcctc catatgctgc aagtgcagcc
ctaaaaaaaa 1500 taaaaaaata aaaaaaaata aataaaagaa gtagacaaat
tgtatagaac aaccctgagt 1560 atgttgcctg agcacatata acaagggtaa
gtattatttc aggaaactct ggtttcacag 1620 atactcttgg catatggacc
cctagagtcc tgatgtaaaa tatattcttc ctgggatctt 1680 aggcaagaag
tttgaaagct ccaactctgc actgctgcca aagaaatgat ttttaagtgc 1740
aaaactcttc ccgttccctt ccctgtataa aattccatag gatctctcca gtgcctctag
1800 gataaaggca gttttcattc tctagttcaa ggtgagagaa gattttaatt
atttcacgtt 1860 ttagtgggga attcaagagt ctggcacctg acatttgctg
aactctctcc attatccctc 1920 tctagttccc cagacgcatc ctatggtaga
aattcgcaaa ctagagtgag cgtcagagta 1980 acccaaggaa actgggtaaa
tgcagctccc tgggctctac cccctgagat tctgattcag 2040 tagatctgaa
gcagagccct ggaatatgca tatgcatcat tgtgtcacac caagcattct 2100
gggtaatgag agttgatgtt aggttctcag tagtaagaca agtatagaga ttccggggga
2160 ctgagtgctc agctctgcct tggggaggag ggagagggct aaagagaaca
ggagatgggg 2220 acagggaatg ctcaacctcc aatcttaggc atttgagcta
tgtcttaggg gtcaggagga 2280 ggttaccaat atagtgatta agagattgag
gttccagtca gagggatatg ctggagaagg 2340 ggggtgaaaa taatgtcata
ggtttggtga gtgcagatac tttgagtttt ttaatatttt 2400 tattgaaata
tagttgattt acaatgctct tagtgagtac aattactttg aataagtgca 2460
tagatgtatg ccattcttcc agaaatgatt tattgagctc ctttgggcat catgctaagt
2520 acaggggaaa cagctgtgaa gaggtccttc ccttatgaag tcattcatcc
ccttcagtaa 2580 atgaaggtaa aggaaaagga tgagacaggg acgccgtgtt
ggaccagggt cagaaaggcc 2640 ttataagacc ttgcctggag ggcaaggaac
ttgcctgtga gtaaggagag cttgagaaag 2700 cgataaagca aagaaggaac
attactgcat tgtgttttag aaaaaccatg tcctggggaa 2760 gaactcctag
agtcaggggg gccagttggg agactgtgct tttttccagg aggagataag 2820
tgaggctgct ggctgagatg gagcaaggat ttagagaagc agatatgaga ttcatttaga
2880 agttagacat tttaggatct gacacataat ttatcaccaa aaccagtgca
tctctggctt 2940 tgggccacca gttttggaga agtggaatgt agggacctac
cattacctgc caatctttac 3000 tacacagatg cctatttccc tcctcatatt
tcctttctcc agatcacgtc ctattctatt 3060 gccaggactc aagattccac
cttgcatgca gtgatccatc ttcacactgg atggacagct 3120 ctagggatgt
cagagcacac tcttgtccat actgctgact gggtctcctg tcagcccatc 3180
tgtctatcag ctgtggtatt attagtataa taagagggct gtatatgaga gacacaaaat
3240 tctaggtgta gctcaaagat aggctagagt tattcctatg tacaacaaat
atttatggga 3300 ccccttctgt gtactgtcat ggttgctgct ttcatcatac
ttgtagtcta atggaggtgg 3360 gggcagggca ggaataagcg gatgtccaca
aaatcagtaa gaccacttat attcaacatt 3420 ttcataattt agttatttga
gcccaaaggg tccacatccg tggtattcca actttttttt 3480 ccccggacat
ggatctttat cttttttttt ttttcttttt tgcggccaga cctgcggcat 3540
atggaagttc ccaggccagg ggttgaatgg gagttgcagc tgcctggtct acaccacagc
3600 cacagcaagg tgggatctga gctgcatctg tgacatacac cgcagctgag
gtaacaccag 3660 attctgaacc cactgaatga ggccagggat ggaacccgtc
tccttatgaa cactatgtca 3720 tgttcttcac cctctgagcc acaacgggaa
ctccagactt cgtctttaaa tgtattctga 3780 cttggagagc tatcacacta
agcaattaac aggagctgac ctggtttagg ctggggtggg 3840 gccctactcc
tcaatgttcc ctgaggcaca tctgtgggac ccctgggcat catctatctg 3900
agcagcctta gagctgctca tccagttgac tgttgatgta gaagtgcaaa cttctgcctt
3960 ccttatttgt tgctttcttt tttcattgtt ctctcccctt tgtgtcttta ag 4012
41 2281 DNA Porcine 41 gtatgctatg aagttattat ttgtttttgt tttcttgtat
tacagagcta tatgaaaacc 60 tcttagtatt ccagttggtt tctcaataag
cattcattga gccttactga ctgtcagacg 120 gagggcgtat tggactatgt
gctgaaacaa tcctttgttg aaaatgtagg gaatgttgaa 180 aatgtaggga
atgaaatgta gatccagctc tgtttctctt ttggaggatt ctttttcctc 240
catcaccgtg tcttggttct tgtttgtttt gggtttttgt gggtgttgta ttgtgttgtg
300 ttggttatgg cagtgacagc tatttaaact gtgaaacggg ggagttcccg
tcgtggcgca 360 gtggttaacg aatccgactg ggaaccatga ggttgcgggt
tcggtccctg cccttgctca 420 gtgggttaac gatccggcgt tgccgtgagc
tgtggtgtag gttgcagaca cggctcggat 480 cccgcgttgc tgtggctcta
gcgtaggcca gcggctacag ctccgattgg acccctagcc 540 tgggaacctc
catatgccgc aggagcggcc caaagaaata gcaaaaagac aaaataaata 600
aataaataaa taagtaagta aaataaactg tgaaacgggg agttcccttc atggctcagc
660 agttaacaaa cccagctagg atccatgagg atgtaggttc gatccctggc
cttgctcagt 720 gggttaagaa tccagcgttg ctgtgagctg tgatgtaggt
cgcagatgca gcccagatcc 780 tgcattgctg tggctgtggc gtaggctggc
agctgaagct ccgattcaac ccctagcctg 840 ggaacatcca tatgctgcag
gtgtggcctt aagaggcaaa aaaataaaaa aataaaaaat 900 aaataaattg
tgggacagac aggtggctcc actgcagagc tggtgtcctg tagcagcctg 960
gaagcaggta aggtaaggac tgcagctggg taaggactga attgcaccaa ctgggaagta
1020 agcctagatc tagaacttaa gttagccctg acatagacac acagagctca
ccagctaagt 1080 ggttcagctt ataagctggt cactgaaact gaggatgtcc
acaaaagcaa aataagtagc 1140 aacaggcagc gggatgcaag agaaagagga
ggcctaaaat ggtctgggaa tccctgccat 1200 acctatattt tatcctactt
atatttagtg cctgaatgtg tgcctggaga gcaaagttta 1260 gggaaagcat
cgggaaatgc acagtattca tacccttagg aacaaagatc agttacctcc 1320
agggtaaaga ctatttccaa gtttaaattt caacccctga acattagtac tgggtaccag
1380 gcaacacttg ccatcctcaa aatcaatgaa tcctaaaatt caacctgggg
gtcagtgaca 1440 gtctgtgaca aagtttttgc tggtcagtaa cgaaataagt
atgagcacca tctgagtatg 1500 gtcaccaaga tgtcaactct ctttcctttg
gacgaattgt cattattcca agattaggtc 1560 ctttctattt ttgaggtgtg
aaaacatctt tcctttcata aaataaaagg atagtaggtg 1620 gaagaatttt
ttttgttttt tggtcttttt gctatttctt tgggccgctt ctgcagcata 1680
tggaggttcc caggccaggg gtcgaatcgg agctttagcc accggcccac gccagagcca
1740 cagcaacacg ggatccaagc cgcatctgca gcctacacca cagctcacgg
caatgccgga 1800 tcgttaaccc actgagcaag ggcagggacc gaacccgcaa
cctcatggtt cctagtcgga 1860 ttcgttaacc actgcgccac gacgggaact
cctaatgata ctcttttata tttagctact 1920 atgtgatgat gagaaacagt
ccacatttta ttatttttta gccaatttga tatctcatta 1980 ctaagataat
gataattttc tctataaatt ttatttaagt tagtgttatg aagtggtttt 2040
gctagtgtag aaggctagga tttgaattca gttcaagaaa gaagagaggg agggagggag
2100 agggatgggt agagggatgg ggcagtggga gagagcaaag aggagagaca
gtttttgtat 2160 taattctgct tcattgctat catttaaggg cacttgggtc
ttgcacattc tagaattttc 2220 taaggacctt gaccgccaga ttgatatgct
tcttcccttt accatgttgt catttgaaca 2280 g 2281 42 4559 DNA Porcine 42
gtaagcagga ataccagtgg aagtgcccct ttcttccttc cttcctaaat aaactttttt
60 attttggaac aactttagag ttacagaaaa gttgcaaaga tattatagac
agtagtgttt 120 atatatatat ataaattttt ttttgctttt tatgaccaca
cctgtggcat atggaggttc 180 ccagtctagg ggttgaattg gagctacagc
tgccagtctg tgccataacc acagcaatgc 240 aggatctggg ccacgtctgt
gacctacacc aaagctcaca gctggattct taacccactg 300 agcaaggcca
gggattgaac ctgcatcctc gtggttccta gttggattcg tttccgcttt 360
gccgcaatgg gaactccaaa ttattgttaa tatcttactt tactggggta catttgttac
420 aaccaatact ctgatactga aacattactg ttaactccgt acttgcttct
ttttgagtca 480 tttgcaaaga ctggcttctt gacctgcttc cttccaaaca
gctggcctgc ctatgctgtt 540 ctcagacctg caagcactga tctctgcccc
ccttgccttc tctccagtgg tgtctccttc 600 cccaaacaaa cccagtgtgg
ctctggaaag ggagttaagt caacataaac caacacatat 660 tttgttgagc
tccaattttg agcaaatccc tcacctacgg cagacaggca tgatgttaag 720
aactagggct ttggacacaa ggtcaagacc aagaagggtt cctcacccct actgattcag
780 ataaccaata atgaggcttt gaatccctgt ccaaaggttg ttttttttcc
cttctattga 840 gcttcttgcc accttatcag ttttttttat gacagtcaaa
tgacatgata tatgtgagca 900 tacatggtaa tttttaattc tatataaatg
aatcactaaa taaattagga ggatatatag 960 tccaccttta agcgtattac
acgtgtcaca tgaatgtgtg gcgacttaat tgtagaggtt 1020 taaatgtagc
ttcctataat agatgtgttc ctaaactaca ttttaatcat tggacttgta 1080
tttttatgtt agcacttgct gttgaagaaa agcctatgcc aaaagttcag tgaaaccaat
1140 aatccactgc cagctttctg agttaaaaaa aatccctggg ttttcacaca
caggaacacc 1200 ctgtgtgaaa cactcattta gagcaaaatg catctgataa
ggagttcctg ttgtgcctca 1260 actggttaag gacctgacat tctccatgag
aatgtgagtt tgatccccgg ccccactcga 1320 tgggttaagg atctggtgtt
gccacaaact gcagctccga ttcatctcct agcctagaaa 1380 cttccacagc
ccagaatatg ccacagaatt cggctgttta aaaaaaaaaa gaaaaaaaaa 1440
agaatcataa atgtgttggt ttgttcacca aatacatgat aacttgctct tgccaagctc
1500 agcttcataa atattaagtc atttaataca gcagccacct tatgaacaga
tattactata 1560 cttcccattt acagataagg aaaatgccat atttaaccaa
gagattaaat aactttcccg 1620 aggtcttata gcaagtaaat catggtgcag
gggtttgacc acacgcagtc tatctccaga 1680 gtctgtgtat ttagccactg
ttttactttc aaatttaaat ttataaaact tctaaattat 1740 ctgttaacca
taatctttgg aatttttaaa accacgagtt cctataaaat gtttcattga 1800
aagtaagtca cttttccata gcttttgata atacatctgt aggataaagt aagccacagc
1860 tctcttgcag acttggtaca ccctggggca aagcatcatg cctgtcacgt
acatggtggt 1920 ccttactttg actctcagtg cttttattgc ccaggaattt
tgtgagattt ctagttgttg 1980 aggtttgttt aaagaggtta tgccggtact
tggaagagct cttttcttgc tacctggagc 2040 cttctcatat ttcctttttg
aggagggaca tgaattgcct ttcaaactca taaatatatt 2100 ttctagtaca
caagtctcca tcttccttag acgcatggct cctggagttc tccatcctcc 2160
tgctccactt tgggtgggct cctctctggg tctgccacca atctgccacc cagagacatc
2220 cttgacccac ttccagaccc caccatggct tcactttctt cgctttcctc
ctttgtggaa 2280 ccttctgctt aagaatctga ggaagaaaat ttgcacgtga
gctaaactgg aggtactttc 2340 ctgcctggtc ttgcacgata gcttggctga
gcccatgatg ctgggtggct gttactttcc 2400 atggacaccc gaaggcgttg
ctcctttggc ttctagttgc atgcagtgtt gcttatccca 2460 ggctgatctt
tcttccactg taggtgactt ttaagaatta agggattaat ctatatctac 2520
aacaacaaca acaaagacct tttcaagctg aggtagggct ttctgtatat gtttggagtg
2580 gttatccagc agactttact tgaaggcagg ggtcatatcc tcaagtgctc
ataaacggac 2640 cacagaaaga tctcataatt gggtggagct gggtggggac
cgtgtcatgt ggccaggaaa 2700 tgccagatgg gaagggagtg gcccttactg
agctccagct gaactctgaa ttttctagaa 2760 aactcagaaa tctggatttt
tcatgtgtaa tacccagatt tatagatgtg gaaagctaat 2820 tctttttttt
tttaagggac tataggcaat gaactaagat ctaggttgta tttggacaag 2880
gggtcatcag tttaagctgt gtagttgagc gctcagctat tgggctgagg gacccctaaa
2940 tactgagacg gggaggtcct tgctctgggg catcacaagt acactccctg
gtctcattca 3000 aacacttttc ctacaaaatt gatcccattt cttcagtgca
ctgtctgaat gcatttggcc 3060 cagagccgtg ctgaggcata gggaaggggt
ccacggtttc atggcatcgt tttgtgctgt 3120 gtgtccctgc tgtcgtccag
gatacctacc tctcctcctc ctgcatctga atgtcccccc 3180 acagactctc
tgggattcta cagcctctgg cctgttcctc agacacctct tacctgccag 3240
ctttccagat tcacattagt tagtccaaat ctactgccgt cagtgactca cttcatttct
3300 tcttctccga ggcagttcag cccggtacag ttgttttgtc aacacttcag
ttgagtctgg 3360 aagatgtgca tgggttatgc acgagagcgg tccatcattt
tgagctagaa gtcctttctc 3420 agcccagaga caagtcctca tctcctttac
ttcctgactc ttcttcctct gcatccttcc 3480 aagatatctc tttctccagc
caccacctaa atctcttctt ttcccggggt tccgtgctca 3540 acccactctt
cttcttaaat ctgtggctgg gtgaacgcat ctgctggcac cacttctctg 3600
ctaaagactc caaaaatcca taggtcctgc ccggcctttg cccacctctc tccaacactg
3660 tccagcttta gatgtagagc taatcccccc agagatatca ttccctggat
gtctaagtcc 3720 tttggtatct cactttcagc gtgttcaaaa tcctcttaca
actgttcttt ctccttttcc 3780 atcttgatta ttggcaacat gccagccttt
cccctacccc cagcagtgag ccaagctaga 3840 aacaagggct taatcttcaa
tctttccttc tccatcccta aacctaatga gtctccaagc 3900 ccttcccagt
ttacacccta aatgttgctc aaaacatccc ctagttcttc cacgtgctct 3960
cctctatatt gaaaggtcaa gaaaggccat cttccctcca ctgtgaggaa atagatcttg
4020 atactgcccc tgagctgggc agtcctcgac ctgacaaact gtgcagtgtt
tctaaatctc 4080 tactggcaaa atgagagtgc ctttgacctg tgttgcgatc
tcagatcaca gtggatgtaa 4140 ttgttttata ggaatggtga acgaaaaaga
agtaaatccc taatgccaaa ctcctgatca 4200 ttctatgtca tttaatagcc
tgtcatttat gataaagttt cctctactgg cattagcaca 4260 atacttctca
ggaaaaaaaa atatgatgcc agatactgaa aagctcctgg gtaaacatga 4320
acatgggtac cgataaaatg gtgaagccag tccaatctta gagtgacttc ccttcatgct
4380 acttcatgct cttttttttt ttttttttta agaaaaaccc cttttttttt
tctcacacca 4440 gtcacagagg agaccgaggc ttagcaaggt taaggtcaca
tgattagtaa gtgctgggct 4500 gaaactcaaa accatctctg cttgtctcct
aaccctgtgc acctctgact attcaacag 4559 43 458 DNA Porcine 43
gtaagggcct tgaccaccga attaaggtaa tcttgctctg tggcaggcct tgttttcagt
60 attttaagta cactggctca ggtaatcctc acaacagccc caggaggaat
gttctattac 120 ctccactgta tagatgagga acttgaggca cagaatggtt
gccaaggtca cacagctata 180 ttgggggttc atacccagcc atccaactct
gtctgtactc tctgccactc tgcaccccca 240 gctcctgatc cacttcctgt
ttccatccct cgatttctgc tgcactcagg ggcccctctc 300
cccctcggcc tgtgagatct gcttcagtag gcttttctcc ctgactcctc catccctgtc
360 cttacaggca gctgcttctc tccgggacac gaggggtcca tacggacact
ctctactggc 420 tgggttgcgc ctaactcgtg attcctcctc tgtttcag 458 44
2482 DNA Porcine 44 gtaagtccga aaatgcctgt cgtgtgtgcc ttaggctgct
gcggaggagg ccagggctat 60 ataagcagag tcagtgactg actgtgccct
gcagtgttga tggccatgga gattccaccg 120 ttagagcttt tttctttgtt
aaccttgaag gcaaatctgg ttaggaagat aactttcaaa 180 gagtcaccat
ctggacattc atgcccatgt gcttcaatcc tgtatacaag cagtttagag 240
tacagggaag ggaaggacat tatgaaaggg agagggtgtg tttggatcca gcagctccat
300 cctcagaatt tatctgaaga cactgcaaaa ttactaagaa tcactatgac
aagaatgagg 360 atggggtgat atggcaaagt tgtgatcctg gaagaccttc
atctcccatg ttgcccaact 420 ctgaacatga atttggtgaa ctagttggtt
aaggggatga tcctccaagt ttctccctgg 480 ttgagctcca aaaaccatgt
aagtttctca tagcaaaacc gtataggtcc ttagggcttt 540 agttggaata
tttgtgctga aatgctggaa agccccattt gccatttttg tatttgcaaa 600
ataatcatca agaggggaga atgcattctt tcatgaccac tgaccctctg aaaaggtcag
660 gaatttagtc tgaagtaggc aagcctccta ccccgcttct gccatgagct
tgcacgcaca 720 ggcctgtctt gacatttctt ctttatagat ttctttttga
atatcttgaa attgctttaa 780 aaatatttaa agaatgtaga attatataaa
ataaaaagga aataacccca cacctcccac 840 aaaaccctgt ttcctgcctt
tctccaccca ctctccaggg taacacttgg taacagcata 900 gttgtatcac
cccaggccta tttttgagca tatcagcatt tcaagaaatg tattttttct 960
caataaaaca tcccttatag ttgaggaggg gaggttatca ttcctgggtt ttgttttttt
1020 ttttttttta atgtaatcct ggtacatcgg taatttgcat tttttattca
ttaatatctt 1080 tggtatttct agtgttggga cacacaggtc aacctcagtt
tttgggtttt tttttttgtc 1140 tttttgtctt tctagggcca cacctgcagc
atatggacgt tcccaagcta ggagtctaat 1200 cagagctgta gccaccagcc
tacgtcatag ccatagcaac gtcagatcca agccgtgtct 1260 gtgacctaca
agcacagctc atggcaacac cggatcctta accactgaac gaggccaggg 1320
gatcgaacac acatcctcat ggatcctagt catgttcatt aaccactgag tcatgatggg
1380 aactccaact tcaactattt taatgtctgt aaaacattcc atttggaaac
catttcattt 1440 gtaaagcaaa atgaaaacat tttgttcatt ttcaacagag
ttcgtagctg acttctgttc 1500 tggaaaaaag gaaatggagc aaatttgagt
gagaaagatt caaagataac ttttctttta 1560 aaaaaaatta tatcttggaa
acttctgggc tattgattct gaagactatt tttctatata 1620 ctgttttgat
agcaaagttc ataaatgtga aaggatcctg cgatgaatct tgggaagcag 1680
tcatagccca atatatcttt gttgctttta aaatgagatt tagtttacta aatatttttc
1740 tgatcataaa aataacacag atctaccgca gaaaatttgg aaaaaaaaaa
acttttaaat 1800 tcaaaaaaca gttaaaccac aaatgatccc accatccaga
gagcaatttg tactttggtg 1860 tctagttcat ctttcttttt ctgtttacaa
gcacatatac cacaagcatt ttttcaaaaa 1920 atgaaaatgg gataatacta
tacatacgtc tgtacacctg catagttact gaacagtctt 1980 tgatctaccc
tgtaagtttc taacttttca ttatttgaaa tgatgttttg gcaaagaaat 2040
atgtaggtgt gtctcgcaca ctttcataat gatttcttag gataaatttc ttaggataaa
2100 ttcataatga tttcttataa taatccatac tctgccaact gatcttcagg
gaagccaact 2160 cgccttctca gaaataacat ataacccatt tacttgccct
ctcaccaata ctaggtccta 2220 atgtttttgt gtacagattc tatattttta
catacaagaa ttccttaaag caaggcatgt 2280 cacagaaaaa tagaaggaag
acacaattgt catgtttaag gactgcattc tgtaccaaaa 2340 atgctaagtt
aaatgaacat ctgaaacagt acagaaacgc tatctttcag ggaaagctga 2400
gtaccaggta ctgaacagat tttggcaaat acagcaggca tggatgtttc caaaacatgt
2460 ttttctactt tatctcttac ag 2482 45 1335 DNA Porcine 45
gtaaagcatc ctgcaggtct gggagacact cttattctcc agcccatcac actgtgtttg
60 gcatcagaat taagcaggca ctatgcctat cagaaaacct gacttttggg
ggaatgaaag 120 aagctaacat tacaagaatg tctgtgttta aaaataagtc
aataagggag ttcccatcgt 180 ggctcagtgg taacgaaccc tactagtatc
cattgaggac acaggttcaa tatctggcct 240 cactcagtcg gctaaggatc
cagtgatgcc gtgagctgca gtgtaggcca cagacgtggc 300 tcagatctgg
tgctgctgtg gctatggtgt aggccggccc cctgtaactc caattcgacc 360
cctaggctgg gaacctaaaa agaccccaaa aaagtcgctt taatgaatag tgaatacatc
420 cagcccaaag tccacagact ctttggtctg gttgtggcaa acatacagcc
agttaacaaa 480 caagacaaaa attatcctag gtggtcagtg ggggttcaga
gctgaatcct gaacactgga 540 aggaaaacag caaccaaatc caaatactgt
atggttttgc ttatatgtag aatctaaatt 600 caaagcaaat gagcaaacca
attgaaacag ttatggaaga caagcaggtg gttgtcaggg 660 gggagataag
gggaggcagg aaagacctgg gcgagggaga ttaagaggta ccaactttca 720
gttgcaaaac aaatgagtca ccagtatgaa atgtgcaatg tgggaaatac aggccataac
780 tttataatct cttttttttt tttgtctttt ttgccttttc taaggctgct
cccgtggcat 840 atggaggttc ccaggctagg agtccaaaca gagctgtagc
tgccagccta caccagagcc 900 acagcaacac gggaacctta acccgctgag
caaggccagg gatcgaaccc gagtcctcac 960 agatgccagt agggttcatt
aaccactgag ccacgacagg aattccaggg tctgttgtgt 1020 tcttaaaaca
cttccaggag agtgagtggt atgtcataag taaacaataa atgttaacca 1080
caacaagctt atgaaataaa caggaaagcc atatgaccta caatcagtca ttgggagaat
1140 ccacaaaagg ttgagcagag gatcaattcc agctcacact ccagttttag
attctcccct 1200 gccttaaagc atcacagact acataatctg agctgaagaa
taaaaattaa aactcacccc 1260 agtgcaaaac agaaatgaaa aagtattaaa
acgaggttca tactgttgtt cattagcaat 1320 atcttttatt cacag 1335 46
40431 DNA Porcine misc_feature (16625)..(16625) n is a, c, g, or t
46 ctgccagcct aagccacagc cacagcaacg ctgggtctga gccatgtctg
cagcctatgc 60 cagagctccc cgcagcgccg gatgcttaac ccactgagca
aggccaggga ttgaaccctc 120 gtcctcatgg atagcagttg agttgtttcc
acggaactct taggggaact cctgattatt 180 ttttatttaa atttatattt
ctctgacttt ttcgtgtgct catcagccac tgactgtgta 240 tctccattag
tcatggtttg ttaactctgt cattcaaacc ctcttcatcc ttgctacgca 300
gataacatca ttataataaa atcgtgcctg aagaccagtg acgcccccaa gctaagttac
360 tgcttcccct ggggggaaaa agaagcaccg cgcgggcgct gacacgaagt
ccgggcagag 420 gaagacgggg cagaggaaga cgggggagca gtgggagcag
cgggcagggc gcgggaagca 480 ctggggatgt tccgcgttgg caggagggtg
ttgggcgagc tcccggtgat gcagggggga 540 ggagcctttt ccgaagtagc
gggacaagag ccacgggaag gaactgttct gagttcccag 600 tcccgacgtc
ctggcagcgc ccaggcactg ttattggtgc ctcctgtgtc cacgcgcttc 660
ccggccaggc agccctggcg gatcctattt tctgttcccc cgattctggt acctctccct
720 cccgccctcg gtgcgcagcc gtcctcctgc agtgcctgct cctccagggg
cgaaaccgat 780 cagggatcag gccacccgcc tcctgaacat ccctccttag
ttcccacagg tgagaaggct 840 tcgccgctgc tgccgctggc gccggcagcg
ccctccacgc acttcgtagt gggcgcgcgc 900 cctcctgcat tgtttctaaa
agattttttt ttatccgctt atgctatcag ttactgagga 960 agtatttaca
aatctactat tattttgaat ttgccttttt ctccttatag tttatcagta 1020
tctcttgaga ctgttattgg tgcctgcaaa tttaaaatga ttggggtttt atgaggaagt
1080 gaacctttta tctttatgaa acgcctaact gaggcaatgt taattgctta
aaatactttc 1140 tttattatca gtgtggccat gccagtgtcc tcttggttag
aatttgcctg atctgccaaa 1200 gctgggagat gggggaaagt agagtgggtt
attgaaactg aatatagagt tcagcatcta 1260 aaagcgaggt agtagaggag
gaagctgtgt caacggaaat actgagctgg gttcacatcc 1320 tctttctcca
cacagtctaa tgccttgtgg aagcaaatga gccacagaag ctgaaggaaa 1380
aaccaccatt ctttcttaat acctggagag aggcaacgac agactatgag caggcaagtg
1440 agagggggct ttagctgtca gggaaggcgg agataaaccc ttgatgggta
ggatggccat 1500 tgaaaggagg ggagaaattt gccccagcag gtagccacca
agcttgggga cttggaggga 1560 gggctttcaa acgtattttc ataaaaaaga
cctgtggagc tgtcaatgct cagggattct 1620 ctcttaaaat ctaacagtat
taatctgcta aaacatttgc cttttcatag catcgaacaa 1680 acgacggaga
tcctgttgtg cctctcacct gccgaagctg ccaatctcaa ggaaggaatc 1740
aattttgttc gaaataagag cactggcaag gattacatct tatttaagaa taagagccgc
1800 ctgaaggcat gtaagaacat gtgcaagcac caaggaggcc tcttcattaa
agacattgag 1860 gatctaaatg gaaggtactg agaatccttt gctttctccc
tggcgatcct ttctcccaat 1920 taggtttggc aggaaatgtg ctcattgaga
aattttaaat gatccaatca acatgctatt 1980 tcccccagca catgcctaac
tttttcttaa gctcctttac ggcagctctc tgattttgat 2040 ttatgacctt
gacttaattt cccatcctct ctgaagaact attgtttaaa atgtattcct 2100
agttgataaa cagtgaaact tctaaggcac atgtgtgtgt gtgtgtgtgt gtgtgtgtgt
2160 ttaccagctt ttatattcaa agactcaagc ctcttttgga tttcctttcc
tgctctctca 2220 gaagtgtgtg tgtgaggtga gtgcttgtcc aaacactgcc
ctagaacaga gagactttcc 2280 ctgatgaaaa cccgaaaaat ggcagagctc
tagctgcacc tggcctcaac agcggctctt 2340 ctgatcattt cttggaagaa
cgagtgctgg tacccctttt ccccagcccc ttgattaaac 2400 ctgcatatcg
cttgcctccc catctcagga gcaattctag gagggagggt gggctttctt 2460
ttcaggattg acaaagctac ccagcttgca aaccaggggg atctgggggg ggggtttgca
2520 cctgatgctc ccccactgat aatgaatgag ggattgaccc catcttttca
agctttgctt 2580 cagcctaact tgactctcgt agtgtttcag ccgtttccat
attaggcttg tcttccaccg 2640 tgtcgtgtcg tcaatcttat ttctcaggtc
atctgtgggc agtttagtgc gaatggactc 2700 agaggtaact ggtagctgtc
caagagctcc ctgctctaac tgtatagaag atcaccaccc 2760 aagtctggaa
tcttcttaca ctggcccaca gacttgcatc actgcatact tagcttcagg 2820
gcccagctcc caggttaagt gctgtcatac ctgtagcttg cttggctctg cagatagggt
2880 tgctagatta ggcaaataga gggtgcccag tcaaatttgc atttcagata
aacaacgaat 2940 atatttttag ttagatatgt ttcaggcact gcatgggaca
tacttttggt aggcagccta 3000 ctctggaaga acctcttggt tgtttgctga
cagactgctt ttgagtccct tgcatcttct 3060 gggtggtttc aagttaggga
gacctcagcc ataggttgtt ctgtcaccaa gaagcttctg 3120 caagcacgtg
caggccttga ggtcttccga cttgtggccc ggggactctg ctttttctct 3180
gtcctttttt ctccttagtg ggccatgtcc tgtggtgttg tcttagccag ttgtttaagg
3240 gagtgttgca gctttatgat taagagcatg gtctttcctt gcaaactgct
tggtttagaa 3300 gcctggctcc accacttagc ggctctgtga cctcggacac
atttcttagc ctttctgggc 3360 ctcgctcttc ttcctcataa agtgaaaatg
aaagtagaca aagccttctc tgtctggcta 3420 ctgagaggat ggagtgattt
catacacata aagcacttaa aataatgtct ggcatatgat 3480 acatgctcaa
taaatgtcac ttacatttgc tattattatt actctgccat gatcttgtgt 3540
agcttaagaa cagaggtctt tacaggaatt caggctgttc ttgaatctgg cttgctcagc
3600 ttaatatggt aattgctttg ccacagactg gtcttcctct ccttcaccca
aagccttagg 3660 gggtgaacga tcccagtttc aacctattct gttggcaggc
taacatggag atggcaccat 3720 cttagctctg ctgcaggtgg ggagccagat
tcacccagct ttgctcccag atacagctcc 3780 ccaagcattt atatgctgaa
actccatccc aagagcagtc tacatggtac actcccccat 3840 ccatctctcc
aaatttggct gcttctactt aggctctctg tgcagcaatt cacctgaaat 3900
atctcttcca cgatacagtc aagggcagtg acctacctgt tccaccttcc cttcctcagc
3960 catttttctt ctttgtacat aatcaagatc aggaactctc ataagctgtg
gtcctcattt 4020 tgtcaatcta atttcacagc ctcttggcac atgaagctgt
cctctctctc ctttctgcct 4080 actgcccatg agcagttgtg acactgccac
atttctcctt taacgaccca gcctgctgaa 4140 tagctgcatt tggaatgttt
tcaatttttg ttaatttatt tatttcatct tttttttttt 4200 tttttttttt
tttttttttt agggccgcac ccatgggata tggaggttcc caggctaggg 4260
atccaatggg agctgtagct gctggcctac accacagcca cagcaatgca caattcgagc
4320 caatctttga cctacaccag agctcacggc aacactggat tcttaaccca
ctgattgagg 4380 ccagggatca aactctcgtc ctcatagata cgagtcagat
tcgttaacct ctgagccatg 4440 atagttgtta gttactcatt gatgagaaag
gaagtgtcac aaaatatcct ccataagtcg 4500 aagtttgaat atgttttctg
ccttgttact agaaaagagc attaaaaatt cttgattgga 4560 atgaagcttg
gaaaaaatca gcatagttta ctgatatata agtgaaaata gaccttgtta 4620
gtttaaacca tctgatattt ctggtggaag acatatttgt ctgtaaaaaa aaaaaatctt
4680 gaacctgttt aaaaaaaaaa cttgactgga aacactacca aaatatggga
gttcctactg 4740 ggacacagca gaaatgaatc taactagtat ccatgaggac
acaggtttga tgcctggcct 4800 cgctaagtgg gttaaggata tggtgttgct
gcagctccaa ttcaacccct atcctgggaa 4860 cccccatatg ccaccctaaa
aagcaaaaag aaaggtgctg ccctaaaaag caaaaagaaa 4920 gaaagaaaga
cagccagaca gactaccaaa tatggagagg aaatggaact tttaggccct 4980
atctccaact atcacatccc tatcaccgtc tggtaagaaa tggaaaaaat attactaagc
5040 ctcctttgtt gctacaatta atctgattct cattctgaag cagtgttgcc
agagttaaca 5100 aataaaaatg caaagctggg tagttaaatt tgaattacag
ataaacaaat tttcagtata 5160 tgttcaatat cgtgtaagac gttttaaaat
aattttttat ttatctgaaa tttatatttt 5220 tcctgtattt tatctggcaa
ccatgatcag aaatctttaa acaatcagga agtctttttt 5280 cttagacaaa
tgaaaatttg agttgatctt aggtttagta cactatacta ggggccaagg 5340
gttatagtgt gactattaaa tcacagataa tctttattac tacattattt ccttatactg
5400 gccccacttg gatcttaccc agcttagctt ttgtatgaga gtcatcctta
aagatgactt 5460 tattctttaa aaaaaaaaac aaattttaag ggctgcaccc
atagcatata gaagttccta 5520 ggctagcggt caaattagag ctgcagctgc
cagcctatgc cacagccaca gcaatgccag 5580 atctgagctg catctgtgac
ctacactgca gcttgcagca atgctggatc cttaacccat 5640 tgaacaatgc
cagggattga acacacatcc tcatggatac tgctcaggtt cctaacctgc 5700
tgagccacag ttggaactcc aaagcagact ttattctgat ggctctgctg atctctaaca
5760 cgttattttg tgccatggtg tttatcttca ctttactcaa gtcagggaaa
cacgaagagt 5820 ctcatacagg ataaacccaa ggagaaatgt gcaaagtcac
atacaaatca aactgacaaa 5880 aatcaaatac aaggaaaaaa tatcttcact
ttcaaaatca cctactgatg atgagtttat 5940 atttccttgg atatttgaat
attagctatt tttttccttt catgagtttt gtgttcaacc 6000 aactacagtc
gtttactttg atcacagaat aatgcattta agccttaaat agattaatat 6060
ttattttcac catttcataa acctaagtac aatttccatc caggtctgtt aaatgcacaa
6120 aacacaactg gaagttagat gtaagcagca tgaagtatat caatcctcct
ggaagcttct 6180 gtcaagacga actgggtaaa taccatcaat actgatcaat
gttttctgct gttactgtca 6240 ttggggtccc tcttgtcaac ttgtttccaa
tctcattaga agccttggat gcattctgat 6300 tttaaactga ggtattttaa
aagtaaccat cactgaaaat tctaggcaag ttttctctaa 6360 aaaatccctt
cattcattca tttgttcagt aagtatttga tgagacctta ccatgtgtaa 6420
acattgcact aggtattaag aaatacaaag atggataaga tagagtcggc gtaaatgaga
6480 tgatataatg agacgttata atgaaactca caattccagt tgggaaataa
agtccttcaa 6540 attccatgac tctttctggc acacgttaga ggctacagct
tctgtgtgat tctcatgctg 6600 gctccacttc cactttttcc ttcttcctac
tcaagaaagc ctatagaaat atgagtaaga 6660 agggcttaat cataggaata
aatttgtctc tgttctaagt gattaaaaat gtctttatca 6720 gtataaaaag
ttacttggga agattcttaa aactgctttt acacactgtt ctagaatgac 6780
tgttatataa ataaaaaagt agatttgatc taacacaatt aaatgacctt tggaaatatt
6840 gactaattct caccttgccc ctcaaaggga tgcctgaacc atttccttct
tttgccagaa 6900 agcccccacc ctttgtctgt tgacctagcc taggaaatct
tcagatcacg ttgttagcac 6960 gaactggtta catgtgctgt acaaatacta
tttaattcat ctgattaaaa aaaaagagat 7020 aagaagcaaa agtttgacta
tcttaaactg tttgcgtagg tgagaggaca attgaccatc 7080 tactttatga
gtatgtaacc cagaaactta aagctcctta agggagctaa gtcttttgga 7140
taagacctat agtgagacct tttagcaaaa tggttaagac tgaatggagc tcactagcgt
7200 gggttcatat cctgatgctc aaacacgcaa ttaaatgact ttaggtgggt
tagtctctgt 7260 tccttagttt cctcaatggg agataatatt ggtagtagcg
attttactgg gttgttgaaa 7320 gaacatctgt taaatgttca gaacgtgtta
cgacagagta cagagtaatg atttgcttgt 7380 atatgtatga ctcaaatagt
ctgccatatg ccttgtgact gggtcctgtg gagcaggaag 7440 gagggatttc
ccacccagca gaaagttggg taaactggaa aatagactga ggccaggaaa 7500
tgatgcaaag cgttgatgtt cactgccacg gcaggtgaag ggcagggcca gagttgtcag
7560 tagggtcagg ggaggactgg aaataaccaa gacccactgc acttttcagc
ctttgctcca 7620 gtaaggtaat gttgtgagag tagaaaattt tgttaacaga
acccactttt cagtacagtg 7680 ctaccaatac tgtagtgatt tcataccaca
tcccaagaaa gaaaaagatg gctcaatccc 7740 atgtgagctg agattatttg
gttttattgt taaataaata gcattgtgtg gtcatcatta 7800 aaaaaggtag
atgttaggaa agtagaagga agaagactct cacctacatt ttcatcactg 7860
ttttggtatc tgccagttgt caccttggtc cccttccccg cctctcccct gcctcctctt
7920 cctccttctc ctttttttgg aatacaattc aggtaccata aaatttaccc
ttttagagtg 7980 tttgactcaa tggtttttag tattttcaca tgttgtgcta
ttactatcac tatataattc 8040 caggtcattc acatcacccc ccaaagaaac
cttctaacta ttagcagtcc attcccttct 8100 tccctcagcc cctggcaacc
actaatctac ttactgtctc catggatgtt cctatattga 8160 atcaagctag
cataaacccc acttgctcat ggtcataatt cttttttata gtgctaaatt 8220
acatttgcta atattcaatt aaggatttct atgtccatat tcataaggaa tattggtgtg
8280 tagttttctc tttgtgtgat atctttgtct ggttggggga tcagagtaat
aattactgct 8340 ctcatagaat gaattgagaa gtgttccctc cttttctatt
tattggaaga gtttgtgaag 8400 tatattggta ttgattcttc tttaaacatt
tggtcagatt caccagtgaa gccatctggg 8460 ccatggctaa tctttgtgaa
aagttttttg attactaatt aaatctcttt aatttgttat 8520 gggtctgctc
ctcagacgtt ctagttcttc ttgagtcagt tttgttcatt tgtttcttcc 8580
taggactttc tccctttcat ttggattatt tagattgata gtaatatccc ccttttaatt
8640 cctggctgta gtaatttggg tcttttctct tttttcttgg tcagtttagc
taaaggtttg 8700 taattgtatt aatcttttca aataactaac ttttttgttt
tgtttgtttt ttgttttttg 8760 ttttttgttt tttgtttttt tttgcttttt
aaggctgcac ctgaggcata tggaagttct 8820 caggctagag gtctaatcgg
agctacagct gctggcctat accacaacca tagcaatgcc 8880 agattcaagc
tgcatctgcg acctacacca caactcggcc agggatcaca cccgcaacct 8940
catggttcct agtcggattt gttaaccact gtgccacgac gggaactccc gcccattttt
9000 tttaacacct catactttaa cataaagatg ggcttcacat ggactgatag
ctcaaatgag 9060 gaaggtaaga ctatgaaagt aatggaagaa atgtagacta
tttttgtgac ctagagatta 9120 ctgatacttc ttgacttttc aaacaatact
tcaaaagtac agcccaaagg gaaaaaagaa 9180 agaaaaaaga aacacacata
tacacaaacc tagtgaataa gatatcatcg atacactaca 9240 gatttctatg
aactggaaga ccccatggac aaagttaaag aacatatgat agtttgagtg 9300
attattttgc aatatttaca accaatgagg gaatattatc cagcttatag gaggaagtaa
9360 tgcaaatcga caagaaaaag ataggaaacc caatataaaa attaagaaaa
tacaaaaatt 9420 aagaaaggat atgaactagc attttacaaa agaaaaatct
ccaaaagtca atcagcacat 9480 gaaaatatgc tcaaacctat taattattag
aaaactacag actgaagcaa tgaggtgctt 9540 tactttacat ctttttgact
gataaaaagt tagaaacaaa ggtgatatca aatgtcaggg 9600 ataaaaggat
atagaaatcg tcatgcctgt ggtgggagta tggccggtgc agtcatgtgg 9660
gaaggtaatc tgacagtggt taggcagagc aggtttatga atacactgtg gcccatcaat
9720 cccacgcctg tttatgtacc aaagaaatcc tgttgtggca gaatctatgg
gtccacccct 9780 gggagcatga attaataaaa tgtggcacca gggtgtgtga
aactccagct agagatgaga 9840 tgtccacatg gcaacatgaa tgcatcttag
aaacatagat ttgagtgaaa aagagtaaga 9900 aacagccggg aaacccaata
ccatttataa aaattaaaga tgcacacata caatgtagta 9960 aatattttgc
atgaactttc aaatggttgc ctacaggggg ggagagtaaa gaagagtaga 10020
aaacaaagat aaagggagta agtaagtagc tctgcctgga ctgaatataa tgtgtcatga
10080 actgagaaat atggttaaca taatcctctt aacttgaggt cctaaatgaa
tgaatgagtc 10140 cactattcat ttacccattc tttaatgtgt attgcattat
aatccatttt tttagaacca 10200 acgaattttg ttcccataac tactaatcag
cctgcctttt ctccctcatt cccttatcag 10260 ctcaggggca ttcctagttt
ttcaaacgtt cctcatttga accaaaaata gcatcattgt 10320 ttaaattata
cttgttttca aatacgatgc ttatatattc caagtgtgtt tgcccatttt 10380
cttaggtggt agaaattttt cattctactt ttctatctac tcagattttc ccgttggaat
10440 tatttccatt gctattaaac ttagaagtcc cccctgtgat atgccatttt
tttcatactt 10500 tttaagcact tggttgcttt tctttgtgtc tttaagcacc
tagaatactt ataaccattg 10560 cacagcactg tgtatcaggc agcccttcct
cttccactaa tttatggtcc ttctcttaga 10620 ctatattaaa ctgttattta
attaggatcc tctcttcgtc cttatgattt aattattata 10680 gttttctaat
atgtttttat tataattcct cttcattatt cctccctatt aaaaatttta 10740
atgaattcca tttgtttgtt cttctagtta aatattaagt cataatccaa ataacttaga
10800 tgtcattagt ttatgtggtc aaagtaagga taccacatct ttatagatgc
aggcagttgg 10860 cagatgtcat
gattttcttc agtgcataaa tgcaatttat ctttgagcaa ggggcataaa 10920
aacttttatg gtattggctt tgaaataata gttaagaact gcagactcag tttttcctgc
10980 ttttcttgaa aaagaacact tctaaagaag gaaaatcctt aagcatggat
atcgatgtaa 11040 ttttctgaaa gtctcctgta attccttggg atttttgttg
ttgtttgttg gtcggttttt 11100 ttgggttttt gtttgtttgt tttgttttgt
tttgttttgc ttttagggct gcacctgtgg 11160 catatggaag ttcccaggct
aggggtccaa ctggagctac agctgccagc ctactccaca 11220 gccacagcaa
catgggatcc tagctgcatc tgtgacctaa ccacagctct tggtaatgcc 11280
agattgttaa cccactgagc aatgccagag atcgaatctg cctcctcatg gacactagtc
11340 agattagttt ctgctgagcc acaatgggaa ttcccaattc cttgtatttt
tgaactggtt 11400 atgtgctagc atataatttt gtttcttgaa tctttgtggg
tttttttttt tttttttttt 11460 tgtctcttgt ctttttaagg ctgcacccac
agcatatgga ggttcccagg ctagaggtca 11520 aattggagct acagctgcca
gcctacacaa caactgcagc aaagtggggc ccaacttata 11580 tgacagttcg
tggcaatgcc ggattcctaa cccactgagc agggccaggg atcgaacctg 11640
agtttccagt cagtttcgtt aaccactgag ccatgatagt aactcctgtt tgttcagtct
11700 tgaacctcct ttttaattct ttattccttg agggtgaaat aattgccata
ataatactat 11760 catttattac atgccttctc tgtgctaggc atagtgacac
tttaggattt attatatcac 11820 ttaatcccta caacaactct gcaaagtatg
tatcataatc ctatttgaca gatcaggaaa 11880 ttgcagccca ggatgcagat
aatatgcatc catcacaagt gactagatat agtccctctg 11940 ctattcagca
gggtctcatt gcctttccat tccaaatgca atagtttgca tctattgtat 12000
atgtgttttg gggttttttt gtcttttttt ttttttttgt cttttctggg gcctcaccct
12060 tggcataggt aggttcccag gctaggggtc aaattgaagc tgcagctgcc
agcctacacc 12120 acagccacag caactcggga tctgagcctc atctgcaacc
tacaccaaag ctcacggcaa 12180 caccggatcc ttaacccact gagtgaggcc
agagatcaaa ccggcaacct catggttcct 12240 agtcggattc attaaccact
gagccacgat gggaactccc taaatgcaat agtttgctct 12300 attaacccca
aactcccagt ccatcccact ccctcctcct ccctcttggc aaccacaagt 12360
ctgttctcca tgtccatgat tttcttttct ggggaaagtt tcatttgtgc catttttcat
12420 tttacgggta atttttactt cagtttcttc cactagcagt tgtcttaaag
tgagtataat 12480 taatattcat ttggaaaatg taagcaaaac attttttaaa
gggccatgcc cacagcatat 12540 gaaagtttct gggccagggg ttgaatccag
gctccaagtt gcagctgtgc cctacactgc 12600 agctgggcaa tgctggatcc
tttaacccac tgtgcccggc tagggatcaa acctgcattt 12660 ccacagctac
ccgagccatt gcagttggat tcttaaccca ctgcactaca gtgggaactc 12720
ccacaaaaca ttttttaatg tcctttgaat aaagtaggaa agtgctcgtc tttgagggca
12780 gggcggcaat gccatttcca caaggtttgc tttggcttgg gacctcatct
gctgtcattt 12840 agtaatgaat aaaattgctg acagtaatag gattaactgt
gtgtggagat agccagggtt 12900 agagataaaa acactggaga agtcaaataa
gttgctcgag gtcctctagc taataagcta 12960 ttaagtggga gagtgagggc
tagaaacagg ccatctgtct cccaagcaca tgtccattag 13020 tggtttgctg
atagccttcc agaacaacag agaggactct caaacatggt cttgcctccc 13080
tccaattgat cccctccatg tgcctcacag cgggtctttc taaaattaag ttctgatttt
13140 aattctccct tgctatagca cttaggtatg gctttcagcc gtgcaataaa
aagcaggcaa 13200 gagtggctca atcatatagg aggttgtttt tcttagatcc
caagcaggta atcctgggca 13260 ttatggttgt tctgcgttta tcaaggagcc
aaattctcta tcacctcctg ttctatcctc 13320 ctcagtatct ggctctattc
ttcagcatct caagatggct tgtgctcctc caagcatggc 13380 agtcaaattc
cacacaagag ggggaaatat gaagggcaga cagtgctggt ctcctgagct 13440
gtccctcttt gtcggggaaa taaatgtatt ccttcaagtc ccgtgagact tctgaagtag
13500 acgtctgctt acgtctcacc caccagaact atgtaaactg cacatagtgc
taggtctaca 13560 tagccactca taactgccag ggggtgggaa atctttaaat
aggtgtacca ccacacaatt 13620 aggatgctaa tagtaaggga gaaggagaga
ataggttttg cgcaagccac cagcatgcct 13680 gccacaattg cttaaaattc
ttcattgacc cctcattgcc acaggatgaa atccaaacgc 13740 cttcttagtt
gggaatctga cctacctgtc tctcccacct ggttcagaca ccattctcct 13800
tggtcataaa attccagtca tttgtgaaca tccagctccc ccatgcctcc atgcctttgc
13860 acatgctgtt cttttatctt ttatgttgtc cttttatctt ttatccaaaa
gagatatccc 13920 atcatcacat ctcttttgtc agcccccaaa tactttgtct
ttcaagttca gctggaggat 13980 tacctcctat ttgaaatcag ctttgtctct
tacaaccaaa caaggttttc cttccgagac 14040 actcccacag caccttgaac
tcatctctat caatcattca tttgattgta atgaagttgt 14100 tggtggtatg
cctgtgtctc tgacacatct gcgatctcat gagttcctta agtggaatgt 14160
gaatagcggg atgaacagta ttggtcttca gccctcatct ctgcagatgt tgcttgaccc
14220 aaatgagcgt tgccttttat tttgattttg ctttgatttg tctactccat
gtacttgagc 14280 catgcatttc tgtcttagcg atgcttttta aaagtcattt
tttggttgat tatccagatt 14340 tgtccacctt tgcttctagt tgtagaaaag
gatgaagaaa atggagtttt gcttctagaa 14400 ctaaatcctc ctaacccgtg
ggattcagaa cccagatctc ctgaagattt ggcatttggg 14460 gaagtgcagg
taaggaaatg ttaaattgca atattcttaa aaacacaaat aaagctaaca 14520
tatcaattta tatatatata tatatatata tttttttttt tttttacatc ttatattacc
14580 ttgagtattc ttggaagtgg ctagttagga catataataa agttattctg
aagtcttttt 14640 ttttcttttt ccatggtgag cagtggcttg atgtggatct
cagctcccag acgaggcact 14700 gaacctgagc cgcagtggtg aaagcaccaa
gttctagcca ctagaccacc agggaactcc 14760 ctattctaaa ttcttgagca
cattatttag gaacctcagg aacttggcag gattacagga 14820 aatatatcta
gatttaaaaa aaaatctttt aacagaggtc ccaaaggaga gtcatgcaca 14880
gctatgggag gaagttcaga aactgccctt gctaccagat cactgtcaga taaaatggcc
14940 agctacatgt ttctgcacat tgccctaaga tctttacaaa cttttctgtg
catttttcca 15000 cttttaaaag aaaatttcgg ggttcctgtt gttgctcagt
ggttaacgaa cccaactagt 15060 atccatgggg acaggggttc gagccctggc
ctcactcagt gggttaagaa tctggcattg 15120 ctgtggctgt ggcgtaggct
ggcggctaca gctcagattg gacccctagc ctgagaacct 15180 ccatatgccg
caggtatggc cctaaaaaaa aaaaaaaaga gagagagaga atttcctcca 15240
gaaaaaacac tttggtagtt tgggagaagt aaacaaccaa aaattaattt ttctggagta
15300 ttcgggaagc ttgtaaaaat gggctcttac ttttttgagg agacaaatgg
gaacctaccc 15360 agaagaggca caatcacctg catttgattt cttgacctct
ccctaccttc tttgctggct 15420 ttccacattt ggatttctgt gaccttatct
ctgctccttg gtgttttcat ttttcctgtg 15480 gacgtgccag actatgggaa
gggagtaagg cgttgattta gaatcctgta gtctctgcct 15540 gtctctagtc
attgttttca cccttctcaa aggaccttga catcctgagt gagtccgcaa 15600
gtaatttagg ggagaagcct tagaagccag tgcagccagg ctacatgact gtgtccaccc
15660 actggaacca gtcattttta tacctattca cagcccccct accatttaaa
tccccagagg 15720 tctgccataa catctgtaac tccctttcct ggtaaattgt
gttctaaaag actggtaaca 15780 aaagatattc tgtggtacag agcataatta
aatacctggg agctgatttg agtggggtaa 15840 atcaactggt ttgaccccta
aaacccacca tgagcatttc tgttctaata aagtaatgcc 15900 cgtgctggga
attgtgttct acggaaatgc tcctgctgtg tctttcttga gtcctgtgtc 15960
attgaacatg cttaggagca aaggtccccc atgtggcttg tctgctaacc agcccagttc
16020 cttgttctgg ctggtaatga tccgatcatc tgaatctcac tgtcttccaa
cagatcacgt 16080 accttactca cgcctgcatg gacctcaagc tgggggacaa
gagaatggtg ttcgaccctt 16140 ggttaatcgg tcctgctttt gcgcgaggat
ggtggttact acacgagcct ccatctgatt 16200 ggctggagag gctgagccgc
gcagacttaa tttacatcag tcacatgcac tcagaccacc 16260 tgaggtaagg
aagggtgagc cctcaactcc gaagaaaatg ctgcaataaa agcactgttg 16320
gttttcagct ttttttgtaa tcactgctca ttctgaggta gattcgcttg ggctgataaa
16380 aagagaacta attcagataa atgcttgcat ttgcatagcc tcttttttta
aaaacttttt 16440 tttttttttt ttttttttgg cttttcaggg ctgaacctgt
ggcatatgga ggttcccagg 16500 ctaggggtcg aatcagagct gtagccccgg
gcctatgcca ctgccatagc aacatgcata 16560 gcctcctttt taaagtgcct
tcctgtttta taccattggg atgtgagaag agctattgtg 16620 gaaangagca
tggggtnata accctggacc tctcacgtcc taccctcagg ntagtgggaa 16680
aactctgagt ttaaggacat caaagtgact cctttttagt tacattatgg nggaatcagc
16740 ncatattttt acaaggggcg gagngtaanc tgttggagtt tacaagacat
atggtggcat 16800 tgcaactact taaccctact attatagcac aaaagcagcc
atagtcggtc ctgaaggagc 16860 ctgatgcctt cagctttata ggcaatgacg
tgtgaatatc acaaacagtt tcctgtgtca 16920 ccaaacatga ttgccttttg
atttcccttt caacccttta aaaaaaggta aaagcccttc 16980 ttagcattca
gcagcaggtc gctgtgtttt gccaactcct gatctgtagc atttcgacaa 17040
cactgagctc tcaacttttg aaccctgagt ccaccacatc cttcagtgaa accagagcca
17100 tgtgatacta aggatagaaa cggaaacttc ctgaatccag gcgatcaaat
aggagggaga 17160 aagaggaact ttcattgaca aaaccacaaa tattgtgaat
ggactgttac aaatattgtg 17220 aatgctccta ttcccaaccc cctggcttca
ttacagggtc ctatgtgttc atccttattg 17280 agaaatttgt attgctactg
ccaggttgcc aatacccagc ggtgcccatg gtgttctaaa 17340 atgaagcaat
ttcaacttta tttttttttc ctgtgacttt acatgacaag ttcacatgaa 17400
ggatatactt tgatagtaat gtccatggtt agggaatata cattgtttgc tggttgactg
17460 gcccctggat ttttctattg aaagtccatg agatctcgaa ggcacaggtg
tgttctctcg 17520 ctttttaagg aaagggttta aaaacttaag taattaacag
ctttagtaac aaattaccta 17580 taacacactt aaaaaccgaa taccacccac
tggagtattg tgctacgatt aaaaatctac 17640 ttgtctacta catgatatct
ttgtcccaca gaaggttctg gaaccaaact tgtaatttca 17700 ggattatgag
agccctgagt tcacgcattg tgtaataact atgttgtgtg gtagtcaatt 17760
tgtacagctt gcttagagag aacaatgtca agttaaggag gcgattgctt tatagtgcct
17820 gtcacaagat gccattgcca ttgtcctagc aagagatatt ctatgggagt
atactacatt 17880 ttagtgagga taagaacttt ttatggcatt tagtccggtc
atttcccaac cactgtcctg 17940 aaaaccaatt tcattttgat ttcaggggct
tgtgtgggca aagttgccag gcattaaaaa 18000 gccacttctc aactgtagta
tcacaatgct ttagttgggt agtgtattgc agatagctta 18060 tggctgaaaa
gttaccaagc cttgcagttt tcactccttt gagtttattt ccttgacaga 18120
attgaccctg agttttttga ctcttacctg ctcaactaat aaacaccaga gtcatttatc
18180 tccattgctc ttgtctgacc tttatttacc gaataatgcc ttatgggttc
acaaaaacaa 18240 ggggggaggg ggccagcatg ccttagaaac tgtctttagt
caagaaatgn gattttatta 18300 tgtaaatata tgagtattat aatagatagt
gttattaata gacaccagca agaattgtca 18360 ataatttaaa aatcacaaat
taaaatacat ccatgttagn atcatttatc ctaactccca 18420 aagcccttta
aagtggaaga tttagatgtt aacccagaga ttaaagacat gttcaaagaa 18480
tccttgattt ttttttgaat cccttgtttt tagagaagaa aacctaatga ttttccccct
18540 ctggattcta catattaaat atagttttgg aacttgaata ttagtatggt
taataagtgc 18600 tgatatgctg attttgttta tatttttctt atgagtaaat
atcctatatc accagacatt 18660 atagtctatg tacaaatatg attcttaaac
ctgatagcac attcattaga gttggaattg 18720 cctttttttt tttttttttt
acagttgcac ctgcaacata tgaaagttcc caggctaggg 18780 gttgaatcca
agctgcagct gccaccctac attacagccg tagtaacagc agatccgagc 18840
tgcatctgca acctatgctg cagctcaggg caatgccaga tccactgagt gaagccaggg
18900 atggaacttg catcctcata gagacaacgt cgtgtcctta acccactgag
ccagaacagg 18960 aactccagaa tttcctttca atagaagaag caccaagttt
aggatcagaa agcctgaatt 19020 tgaataccaa tttactattt gttagtcata
tatttctgag tgtgtttcct catttattaa 19080 aagcagacta aaagatgaga
gggtcttttg ttgagaatca aatacaataa catgtgaaag 19140 tgtgtaacac
tatgattgaa atatacctac acagccattt atttgtttat tgttcatgtt 19200
ttgccaccca cacagtagta tataatcctt ttatgtaata aatgctaata atgaaagttg
19260 gcaacttatg taagtactca aaatgctgga ggtcatggga tactgactgg
gatactacag 19320 aggtaatgtc atttcctctg cgctaaactt attgtcttgt
agttagggac tgactctctt 19380 taggacaagg agttcattct gtataccatg
tgtggctatc acccttcgaa gttgaaaaac 19440 tgccccaggg tgggcaccca
tccgttctct tagatatatg gccgagacct ttctctcact 19500 gggagggaac
cacactgagg aatgagaaaa aaaaaaggaa aatcaagatg aaaccagaaa 19560
cctctttggc ataacttctc cactctgtac tttttgttag aactaccctt gcacaaagca
19620 gcatcagtgt ggaagacaga atttgcacac ctggtttgat atacatgccg
tggtatatgg 19680 gatgttctaa caataaagag gactctccca ggaaatctcc
tcactgttat agtcagcctt 19740 gaggaaagag ctcttctttt ggactctggg
gagagtctag tttttcagtt ccttgcttct 19800 cggtcaacgt gttggtgtaa
ggatcacact ctctcttata ctagataatt ctattttttc 19860 acctttcaac
ctgtctatcc ttctgaccct agttacccaa cactgaagaa gcttgctgag 19920
agaagaccag atgttcccat ttatgttggc aacacggaaa gacctgtatt ttggaatctg
19980 aatcagagtg gcgtccagtt gactaatatc aatgtagtgc catttggaat
atggcagcag 20040 gtctgtgttc tttccacatg tttgggttat cctttctggg
ataaatttga ggcgagatag 20100 aaactttaag actaaagaaa caatggccta
ctttttttgt acatggtcct gtgtaaatct 20160 ctatttgagc tgaaataaga
tggtcttcct ctccaattat ccatggtatg actctgatgg 20220 ataacaaatc
cagttctgaa aaaaggggat ttctttccag aagagaggac agtttcttca 20280
aatattgaat taaaagcaaa atagatgtaa accgttgttg gttttattgt tgaattccag
20340 gtagacaaaa atcttcgatt catgatcttg atggatggcg ttcatcctga
gatggacact 20400 tgcattattg tggaatacaa aggtattttc ttgccctcat
cagcatgaaa ttgctcttgg 20460 tagaaaggat aataatagtt atccaaaaca
tcatcctatg ttcatctgtt tcttccctct 20520 tcattttcca tagagtacag
tatattctat ctctgtctta ggaaaatgga ctgtcattca 20580 tataatctta
cagagaatca attagtaatg tactctatgc cgtgacaggt gcgaaggttt 20640
tttttgaagg caacagataa aaatatccta tatttcacct attgtaattt ccttaaaact
20700 gacattattg aataaatgtt ttactttcat cttgaatatt attatgttat
ggaatcatac 20760 actttacccc aataatcatc gaaaagaatt tccaaaaggt
tgagagagtt gtgttgatct 20820 gattactttc ctctgcatcc tttgagctta
acctttgaat atagtttgct aaggaaagta 20880 gtctgtttat gatcctggag
tggaatcagg ctaagtgtcc tcattcagaa cccactgaat 20940 cagacagaat
gaatttattt ccttgaaagt tcaaaatgtg tcactcaaga gtataaattt 21000
tcaaatctta ctctctcttt tccttggatg tgagcaattc ttcgataatt gaatgaggca
21060 gattatatag acttacatgg aagactgttg gcctgagaat tcaaactatg
gtgttcaaga 21120 cttcacngng agtccgatgc catttgtttc ccacaggtca
taaaatactc aatacagtgg 21180 attgcaccag acccaatgga ggaaggctgc
ctatgaaggt tgcattaatg atgagtgatt 21240 ttgctggagg agcttcaggc
tttccaatga ctttcagtgg tggaaaattt actggtaatt 21300 ctttatatca
aaatgatgcc aaggagttgg catggcactt tgctaaatgc tgtgtgaatc 21360
aatacaaaga taattaggac atggttcttc ctcacaagag gtgtgcaatc ttattgggaa
21420 atcatacttg caagtcacaa atatagacta aagtttccag ctgagaatat
gctgatggag 21480 catgaaacac taaggagaca gggagaatct caggaaaaat
caagaataat ttggatcaaa 21540 tggattcctg acatagaaca tagagctgat
cagaaagagt ctgacattgg taatccaggc 21600 ttaagtgctc tttgtatgtg
gttcagaaca gagtgtgggc agcctgaggg ggatacatac 21660 ccttgacctc
gtggaaagct catacggggg agggatgagg ctaaggaagc ccctctaaag 21720
tgtgggatta cgagaggttg ggggggtggt agggaaaata gtggtcaaag agtataaact
21780 tccagttaca agatgaataa attctagggg tataataaca gcatggcact
atagatagca 21840 tattgtacta tatactggaa gtgctgagag tagatcttac
atgttctaac cacacacaca 21900 cacacacaca cacacacacc acacacacac
accacacaca cacacgtgca cacaaacaga 21960 aatggtaatt atgtgaggtg
atggcggtgt taactaactt tattgtggtc atcatttagc 22020 catacatgca
tgtcatgaaa tcaccatgtt gtacacctta aagttatgta atactagatg 22080
tcagttatat ctcaaagcta gaaaaaatgt ggggaccaag gcagaagctc ttctgctctg
22140 tgtctaaggg tggttctggg gctgggatgg ggaggatggt taagtggtat
atttttttca 22200 tacctttgct cagtactatc attgtaagtg ttcaatatat
gtctgcttaa taaattaatg 22260 tttttagtaa gtaatctctg tttagtaatg
tgtcagaaat gccctacttg caataggaag 22320 aaaacctgtc cagtcccttc
cttttttctg taagtctgat ttcattgcct cccagaatgc 22380 atcaccatgt
gagagataga gggaaggtgc tgtccttatg gggttaacag tgtgactagg 22440
gaggcaaaat atacctacta aagggtggta gcataattca gttcttatgt gagtatgtgt
22500 atgtgtgtga gtatgtgcac atgcacatac attttaaaag gtctgtaata
tactaacatg 22560 ttcatagtgg ttacacctag cttataggta acattttttc
ccctgtatcc ttgtttgtgt 22620 ttatcaaatt ttcataacag taatggtaga
aggagtacct gacatggtac catacatgct 22680 nggncctgcc taatttctcn
atttccttta ttgcccatac ccccattgct tgacaagcat 22740 aagtccatac
tggcttgttt tcgttcctca gactcagtac accatgtagc tccatgccct 22800
gggtctttgt atgtgctatt tctactgctt agagtgctat tgcccctgac caccacgtgg
22860 tcagcaactt ctcttctgcg tctgtgtcta tggtctatga ttccagatgt
catcttcact 22920 aactaccctt ctaatatgcc cttccatccc acccgtcctc
atccttaccc cagccactct 22980 ctatttggtg gctctgtttt attttcttcc
tagctcatca ctctttgaaa tgaacttatt 23040 tacttattca atatttgctt
ctttcactag aatgaatgct ccatgagagc agggacctgc 23100 tttatcttgc
tcgccactgt attcacagtg cctagaacta cgtctggcac atagtaggtg 23160
ctcaataaat atcgatcaaa tgaaagaatg agcaaacgaa caaatgaaca acacgtgagg
23220 taggcatcat gattccatca acagaggaga aaaccagact taaagnaatg
aagtggngga 23280 gctgcatttg atcttgactg actccaacat ccatgctctt
gaccactgtg catctccaga 23340 gtgtaatgaa catactttac ttttatattc
caccaaaata acaaagccat gcccatgtta 23400 gtagagagtt aatcgacagt
gcccttaaaa tatgcatgca cccagggtac aactatgcat 23460 gctgccctgt
gttttcagtt ggatccaaat gaattgccgt aaacaaagag gggattcaat 23520
gtctttgact agtttgggat attttcctag taaccaactt tgcaaaataa agccactaat
23580 gacaaggagc tttgttctac ttctgcatca ctcaactgtc aatttttatc
tcttgcaaga 23640 cttctaatct actagaactt ttgtttttct gtgatttctg
aacagagaag actaatccaa 23700 accctgtcat tccagaggaa tggaaagccc
aattcattaa aacagaaagg aagaaactcc 23760 tgaactacaa ggctcggctg
gtgaaggacc tacaacccag aatttactgc ccctttcctg 23820 ggtatttcgt
ggaatcccac ccagcagaca agtatggctg gatattttat ataacgtgtt 23880
tacgcataag ttaatatatg ctgaatgagt gatttagctg tgaaacaaca tgaaatgaga
23940 aagaatgatt agtaggggtc tggagcttat tttaacaagc agcctgaaaa
cagagagtat 24000 gaataaaaaa aattaaatac aagagtgtgc tattaccaat
tatgtataat agtcttgtac 24060 atctaacttc aattccaatc actatatgct
tatactaaaa aacgaagtat agagtcaacc 24120 ttctttgact aacagctctt
ccctagtcag ggacattagc tcaagtatag tctttatttt 24180 tcctggggta
agaaaagaag gattgggaag taggaatgca aagaaataaa aaataattct 24240
gtcattgttc aaataagaat gtcatctgaa aataaactgc cttacatggg aatgctctta
24300 tttgtcaggt atattaagga aacaaacatc aaaaatgacc caaatgaact
caacaatctt 24360 atcaagaaga attctgaggt ggtaacctgg accccaagac
ctggagccac tcttgatctg 24420 ggtaggatgc taaaggaccc aacagacagg
tttgacttga atatttacag ggaacaaaaa 24480 tgatttctga attttttcat
gtttatgaga aaataaaggg catacctatg gcctcttggc 24540 aggtccctgt
ttgtaggaat attaagtttt tcttgactag catcctgagc ttgtcatgca 24600
ttaagatcta cacaccaccc tttaaagtgg gagtcttact gtataaaata aactattaaa
24660 taagtatctt tcaactctgg ggtggggggg gagactgagt tttttcacag
tcctatataa 24720 taattttctt atcctataaa ataattagga gttcccgtag
tggctcagca atagcaaacc 24780 cgactagtat cgatgaggat gcgggttcga
ttcctggccc ccctcagtgg gttaaggatc 24840 tggcattgcc gtgagctgtg
gtgtaggtgg cagacacggc tcagatccca cgttactgtg 24900 gctgtggcat
aggccagcag ctccagctct gattagaccc ttagcctggg aacttccata 24960
tgctgtgggt gtggccttga aaaaaaataa ataaataaga taattactca aatgttttcc
25020 ttgtctcaga accttacttc aggataaaga gtgagaaagt tttttttatg
aagggccatt 25080 attacagctc aaaaataagt tgtcttcagc aagtagaaag
caataagcct gagagttagt 25140 gttcctatca gtgtaaatat tacctcctcg
ccaatcccca gacagtccat ttgaacaatt 25200 aacggtgccc tgggagtaca
gttcagaaac attaatgtgg atgttccaga cctgtatttt 25260 tataagtact
tgtcttgagc cggatggaac catcattcct caccattatt tagaagtgga 25320
ctgtgactct gttggagatc agggcacacg gttaccaaaa gcacaccctt ctcctggcct
25380 tacctttgca aagctggggt ctgggacaca gtcagctgat tatacccttt
tactaacttc 25440 ccacagctca aatctggtca attctccttc acaaatctct
taaaaatcca tcactcacct 25500 ccagcctctt ctgctgtggc cttgattcag
cctctcacaa ttttttttta accagaattc 25560 tggcagtggc ccctgacttg
cctctgtgct cccagccccg ctgtcctctg atccatcctc 25620 catgccagcc
tttttcaatc tgctggtcac gattcattga tgggttagga aatcaatggc 25680
atcacaacta gcatttagaa aaaggaaata ggcgttcccg ccgtggcaca gcagaaataa
25740 atccgactag gaaccataag gttgcgggtt caacccctgg ccttgttcag
tgggttaagg 25800 atccggcatt gccgtgggct gttttgtaag tcacagacat
ggctctgatc cggcattgct 25860 gtggctctgg cgtaggcctg cagcatcagc
tccaattaga cccctatcct gggagcctcc 25920 atatgctgca
agtgcagccc taaaaaaaat aaaaaaataa aaaaaaataa ataaaagaag 25980
tagacaaatt gtatagaaca accctgagta tgttgcctga gcacatataa caagggtaag
26040 tattatttca ggaaactctg gtttcacaga tactcttggc atatggaccc
ctagagtcct 26100 gatgtaaaat atattcttcc tgggatctta ggcaagaagt
ttgaaagctc caactctgca 26160 ctgctgccaa agaaatgatt tttaagtgca
aaactcttcc cgttcccttc cctgtataaa 26220 attccatagg atctctccag
tgcctctagg ataaaggcag ttttcattct ctagttcaag 26280 gtgagagaag
attttaatta tttcacgttt tagtggggaa ttcaagagtc tggcacctga 26340
catttgctga actctctcca ttatccctct ctagttcccc agacgcatcc tatggtagaa
26400 attcgcaaac tagagtgagc gtcagagtaa cccaaggaaa ctgggtaaat
gcagctccct 26460 gggctctacc ccctgagatt ctgattcagt agatctgaag
cagagccctg gaatatgcat 26520 atgcatcatt gtgtcacacc aagcattctg
ggtaatgaga gttgatgtta ggttctcagt 26580 agtaagacaa gtatagagat
tccgggggac tgagtgctca gctctgcctt ggggaggagg 26640 gagagggcta
aagagaacag gagatgggga cagggaatgc tcaacctcca atcttaggca 26700
tttgagctat gtcttagggg tcaggaggag gttaccaata tagtgattaa gagattgagg
26760 ttccagtcag agggatatgc tggagaaggg gggtgaaaat aatgtcatag
gtttggtgag 26820 tgcagatact ttgagttttt taatattttt attgaaatat
agttgattta caatgctctt 26880 agtgagtaca attactttga ataagtgcat
agatgtatgc cattcttcca gaaatgattt 26940 attgagctcc tttgggcatc
atgctaagta caggggaaac agctgtgaag aggtccttcc 27000 cttatgaagt
cattcatccc cttcagtaaa tgaaggtaaa ggaaaaggat gagacaggga 27060
cgccgtgttg gaccagggtc agaaaggcct tataagacct tgcctggagg gcaaggaact
27120 tgcctgtgag taaggagagc ttgagaaagc gataaagcaa agaaggaaca
ttactgcatt 27180 gtgttttaga aaaaccatgt cctggggaag aactcctaga
gtcagggggg ccagttggga 27240 gactgtgctt ttttccagga ggagataagt
gaggctgctg gctgagatgg agcaaggatt 27300 tagagaagca gatatgagat
tcatttagaa gttagacatt ttaggatctg acacataatt 27360 tatcaccaaa
accagtgcat ctctggcttt gggccaccag ttttggagaa gtggaatgta 27420
gggacctacc attacctgcc aatctttact acacagatgc ctatttccct cctcatattt
27480 cctttctcca gatcacgtcc tattctattg ccaggactca agattccacc
ttgcatgcag 27540 tgatccatct tcacactgga tggacagctc tagggatgtc
agagcacact cttgtccata 27600 ctgctgactg ggtctcctgt cagcccatct
gtctatcagc tgtggtatta ttagtataat 27660 aagagggctg tatatgagag
acacaaaatt ctaggtgtag ctcaaagata ggctagagtt 27720 attcctatgt
acaacaaata tttatgggac cccttctgtg tactgtcatg gttgctgctt 27780
tcatcatact tgtagtctaa tggaggtggg ggcagggcag gaataagcgg atgtccacaa
27840 aatcagtaag accacttata ttcaacattt tcataattta gttatttgag
cccaaagggt 27900 ccacatccgt ggtattccaa cttttttttc cccggacatg
gatctttatc tttttttttt 27960 tttctttttt gcggccagac ctgcggcata
tggaagttcc caggccaggg gttgaatggg 28020 agttgcagct gcctggtcta
caccacagcc acagcaaggt gggatctgag ctgcatctgt 28080 gacatacacc
gcagctgagg taacaccaga ttctgaaccc actgaatgag gccagggatg 28140
gaacccgtct ccttatgaac actatgtcat gttcttcacc ctctgagcca caacgggaac
28200 tccagacttc gtctttaaat gtattctgac ttggagagct atcacactaa
gcaattaaca 28260 ggagctgacc tggtttaggc tggggtgggg ccctactcct
caatgttccc tgaggcacat 28320 ctgtgggacc cctgggcatc atctatctga
gcagccttag agctgctcat ccagttgact 28380 gttgatgtag aagtgcaaac
ttctgccttc cttatttgtt gctttctttt ttcattgttc 28440 tctccccttt
gtgtctttaa gcaagggcat cgtagagcct ccagaaggga ctaagattta 28500
caaggattcc tgggattttg gcccatattt gaatatcttg aatgctgcta taggagatga
28560 aatatttcgt cactcatcct ggataaaaga atacttcact tgggctggat
ttaaggatta 28620 taacctggtg gtcagggtat gctatgaagt tattatttgt
ttttgttttc ttgtattaca 28680 gagctatatg aaaacctctt agtattccag
ttggtttctc aataagcatt cattgagcct 28740 tactgactgt cagacggagg
gcgtattgga ctatgtgctg aaacaatcct ttgttgaaaa 28800 tgtagggaat
gttgaaaatg tagggaatga aatgtagatc cagctctgtt tctcttttgg 28860
aggattcttt ttcctccatc accgtgtctt ggttcttgtt tgttttgggt ttttgtgggt
28920 gttgtattgt gttgtgttgg ttatggcagt gacagctatt taaactgtga
aacgggggag 28980 ttcccgtcgt ggcgcagtgg ttaacgaatc cgactgggaa
ccatgaggtt gcgggttcgg 29040 tccctgccct tgctcagtgg gttaacgatc
cggcgttgcc gtgagctgtg gtgtaggttg 29100 cagacacggc tcggatcccg
cgttgctgtg gctctagcgt aggccagcgg ctacagctcc 29160 gattggaccc
ctagcctggg aacctccata tgccgcagga gcggcccaaa gaaatagcaa 29220
aaagacaaaa taaataaata aataaataag taagtaaaat aaactgtgaa acggggagtt
29280 cccttcatgg ctcagcagtt aacaaaccca gctaggatcc atgaggatgt
aggttcgatc 29340 cctggccttg ctcagtgggt taagaatcca gcgttgctgt
gagctgtgat gtaggtcgca 29400 gatgcagccc agatcctgca ttgctgtggc
tgtggcgtag gctggcagct gaagctccga 29460 ttcaacccct agcctgggaa
catccatatg ctgcaggtgt ggccttaaga ggcaaaaaaa 29520 taaaaaaata
aaaaataaat aaattgtggg acagacaggt ggctccactg cagagctggt 29580
gtcctgtagc agcctggaag caggtaaggt aaggactgca gctgggtaag gactgaattg
29640 caccaactgg gaagtaagcc tagatctaga acttaagtta gccctgacat
agacacacag 29700 agctcaccag ctaagtggtt cagcttataa gctggtcact
gaaactgagg atgtccacaa 29760 aagcaaaata agtagcaaca ggcagcggga
tgcaagagaa agaggaggcc taaaatggtc 29820 tgggaatccc tgccatacct
atattttatc ctacttatat ttagtgcctg aatgtgtgcc 29880 tggagagcaa
agtttaggga aagcatcggg aaatgcacag tattcatacc cttaggaaca 29940
aagatcagtt acctccaggg taaagactat ttccaagttt aaatttcaac ccctgaacat
30000 tagtactggg taccaggcaa cacttgccat cctcaaaatc aatgaatcct
aaaattcaac 30060 ctgggggtca gtgacagtct gtgacaaagt ttttgctggt
cagtaacgaa ataagtatga 30120 gcaccatctg agtatggtca ccaagatgtc
aactctcttt cctttggacg aattgtcatt 30180 attccaagat taggtccttt
ctatttttga ggtgtgaaaa catctttcct ttcataaaat 30240 aaaaggatag
taggtggaag aatttttttt gttttttggt ctttttgcta tttctttggg 30300
ccgcttctgc agcatatgga ggttcccagg ccaggggtcg aatcggagct ttagccaccg
30360 gcccacgcca gagccacagc aacacgggat ccaagccgca tctgcagcct
acaccacagc 30420 tcacggcaat gccggatcgt taacccactg agcaagggca
gggaccgaac ccgcaacctc 30480 atggttccta gtcggattcg ttaaccactg
cgccacgacg ggaactccta atgatactct 30540 tttatattta gctactatgt
gatgatgaga aacagtccac attttattat tttttagcca 30600 atttgatatc
tcattactaa gataatgata attttctcta taaattttat ttaagttagt 30660
gttatgaagt ggttttgcta gtgtagaagg ctaggatttg aattcagttc aagaaagaag
30720 agagggaggg agggagaggg atgggtagag ggatggggca gtgggagaga
gcaaagagga 30780 gagacagttt ttgtattaat tctgcttcat tgctatcatt
taagggcact tgggtcttgc 30840 acattctaga attttctaag gaccttgacc
gccagattga tatgcttctt ccctttacca 30900 tgttgtcatt tgaacagatg
attgagacag atgaggactt cagccctttg cctggaggat 30960 atgactattt
ggttgacttt ctggatttat cctttccaaa agaaagacca agccgggaac 31020
atccatatga ggaagtaagc aggaatacca gtggaagtgc ccctttcttc cttccttcct
31080 aaataaactt ttttattttg gaacaacttt agagttacag aaaagttgca
aagatattat 31140 agacagtagt gtttatatat atatataaat ttttttttgc
tttttatgac cacacctgtg 31200 gcatatggag gttcccagtc taggggttga
attggagcta cagctgccag tctgtgccat 31260 aaccacagca atgcaggatc
tgggccacgt ctgtgaccta caccaaagct cacagctgga 31320 ttcttaaccc
actgagcaag gccagggatt gaacctgcat cctcgtggtt cctagttgga 31380
ttcgtttccg ctttgccgca atgggaactc caaattattg ttaatatctt actttactgg
31440 ggtacatttg ttacaaccaa tactctgata ctgaaacatt actgttaact
ccgtacttgc 31500 ttctttttga gtcatttgca aagactggct tcttgacctg
cttccttcca aacagctggc 31560 ctgcctatgc tgttctcaga cctgcaagca
ctgatctctg ccccccttgc cttctctcca 31620 gtggtgtctc cttccccaaa
caaacccagt gtggctctgg aaagggagtt aagtcaacat 31680 aaaccaacac
atattttgtt gagctccaat tttgagcaaa tccctcacct acggcagaca 31740
ggcatgatgt taagaactag ggctttggac acaaggtcaa gaccaagaag ggttcctcac
31800 ccctactgat tcagataacc aataatgagg ctttgaatcc ctgtccaaag
gttgtttttt 31860 ttcccttcta ttgagcttct tgccacctta tcagtttttt
ttatgacagt caaatgacat 31920 gatatatgtg agcatacatg gtaattttta
attctatata aatgaatcac taaataaatt 31980 aggaggatat atagtccacc
tttaagcgta ttacacgtgt cacatgaatg tgtggcgact 32040 taattgtaga
ggtttaaatg tagcttccta taatagatgt gttcctaaac tacattttaa 32100
tcattggact tgtattttta tgttagcact tgctgttgaa gaaaagccta tgccaaaagt
32160 tcagtgaaac caataatcca ctgccagctt tctgagttaa aaaaaatccc
tgggttttca 32220 cacacaggaa caccctgtgt gaaacactca tttagagcaa
aatgcatctg ataaggagtt 32280 cctgttgtgc ctcaactggt taaggacctg
acattctcca tgagaatgtg agtttgatcc 32340 ccggccccac tcgatgggtt
aaggatctgg tgttgccaca aactgcagct ccgattcatc 32400 tcctagccta
gaaacttcca cagcccagaa tatgccacag aattcggctg tttaaaaaaa 32460
aaaagaaaaa aaaaagaatc ataaatgtgt tggtttgttc accaaataca tgataacttg
32520 ctcttgccaa gctcagcttc ataaatatta agtcatttaa tacagcagcc
accttatgaa 32580 cagatattac tatacttccc atttacagat aaggaaaatg
ccatatttaa ccaagagatt 32640 aaataacttt cccgaggtct tatagcaagt
aaatcatggt gcaggggttt gaccacacgc 32700 agtctatctc cagagtctgt
gtatttagcc actgttttac tttcaaattt aaatttataa 32760 aacttctaaa
ttatctgtta accataatct ttggaatttt taaaaccacg agttcctata 32820
aaatgtttca ttgaaagtaa gtcacttttc catagctttt gataatacat ctgtaggata
32880 aagtaagcca cagctctctt gcagacttgg tacaccctgg ggcaaagcat
catgcctgtc 32940 acgtacatgg tggtccttac tttgactctc agtgctttta
ttgcccagga attttgtgag 33000 atttctagtt gttgaggttt gtttaaagag
gttatgccgg tacttggaag agctcttttc 33060 ttgctacctg gagccttctc
atatttcctt tttgaggagg gacatgaatt gcctttcaaa 33120 ctcataaata
tattttctag tacacaagtc tccatcttcc ttagacgcat ggctcctgga 33180
gttctccatc ctcctgctcc actttgggtg ggctcctctc tgggtctgcc accaatctgc
33240 cacccagaga catccttgac ccacttccag accccaccat ggcttcactt
tcttcgcttt 33300 cctcctttgt ggaaccttct gcttaagaat ctgaggaaga
aaatttgcac gtgagctaaa 33360 ctggaggtac tttcctgcct ggtcttgcac
gatagcttgg ctgagcccat gatgctgggt 33420 ggctgttact ttccatggac
acccgaaggc gttgctcctt tggcttctag ttgcatgcag 33480 tgttgcttat
cccaggctga tctttcttcc actgtaggtg acttttaaga attaagggat 33540
taatctatat ctacaacaac aacaacaaag accttttcaa gctgaggtag ggctttctgt
33600 atatgtttgg agtggttatc cagcagactt tacttgaagg caggggtcat
atcctcaagt 33660 gctcataaac ggaccacaga aagatctcat aattgggtgg
agctgggtgg ggaccgtgtc 33720 atgtggccag gaaatgccag atgggaaggg
agtggccctt actgagctcc agctgaactc 33780 tgaattttct agaaaactca
gaaatctgga tttttcatgt gtaataccca gatttataga 33840 tgtggaaagc
taattctttt tttttttaag ggactatagg caatgaacta agatctaggt 33900
tgtatttgga caaggggtca tcagtttaag ctgtgtagtt gagcgctcag ctattgggct
33960 gagggacccc taaatactga gacggggagg tccttgctct ggggcatcac
aagtacactc 34020 cctggtctca ttcaaacact tttcctacaa aattgatccc
atttcttcag tgcactgtct 34080 gaatgcattt ggcccagagc cgtgctgagg
catagggaag gggtccacgg tttcatggca 34140 tcgttttgtg ctgtgtgtcc
ctgctgtcgt ccaggatacc tacctctcct cctcctgcat 34200 ctgaatgtcc
ccccacagac tctctgggat tctacagcct ctggcctgtt cctcagacac 34260
ctcttacctg ccagctttcc agattcacat tagttagtcc aaatctactg ccgtcagtga
34320 ctcacttcat ttcttcttct ccgaggcagt tcagcccggt acagttgttt
tgtcaacact 34380 tcagttgagt ctggaagatg tgcatgggtt atgcacgaga
gcggtccatc attttgagct 34440 agaagtcctt tctcagccca gagacaagtc
ctcatctcct ttacttcctg actcttcttc 34500 ctctgcatcc ttccaagata
tctctttctc cagccaccac ctaaatctct tcttttcccg 34560 gggttccgtg
ctcaacccac tcttcttctt aaatctgtgg ctgggtgaac gcatctgctg 34620
gcaccacttc tctgctaaag actccaaaaa tccataggtc ctgcccggcc tttgcccacc
34680 tctctccaac actgtccagc tttagatgta gagctaatcc ccccagagat
atcattccct 34740 ggatgtctaa gtcctttggt atctcacttt cagcgtgttc
aaaatcctct tacaactgtt 34800 ctttctcctt ttccatcttg attattggca
acatgccagc ctttccccta cccccagcag 34860 tgagccaagc tagaaacaag
ggcttaatct tcaatctttc cttctccatc cctaaaccta 34920 atgagtctcc
aagcccttcc cagtttacac cctaaatgtt gctcaaaaca tcccctagtt 34980
cttccacgtg ctctcctcta tattgaaagg tcaagaaagg ccatcttccc tccactgtga
35040 ggaaatagat cttgatactg cccctgagct gggcagtcct cgacctgaca
aactgtgcag 35100 tgtttctaaa tctctactgg caaaatgaga gtgcctttga
cctgtgttgc gatctcagat 35160 cacagtggat gtaattgttt tataggaatg
gtgaacgaaa aagaagtaaa tccctaatgc 35220 caaactcctg atcattctat
gtcatttaat agcctgtcat ttatgataaa gtttcctcta 35280 ctggcattag
cacaatactt ctcaggaaaa aaaaatatga tgccagatac tgaaaagctc 35340
ctgggtaaac atgaacatgg gtaccgataa aatggtgaag ccagtccaat cttagagtga
35400 cttcccttca tgctacttca tgctcttttt tttttttttt tttaagaaaa
accccttttt 35460 tttttctcac accagtcaca gaggagaccg aggcttagca
aggttaaggt cacatgatta 35520 gtaagtgctg ggctgaaact caaaaccatc
tctgcttgtc tcctaaccct gtgcacctct 35580 gactattcaa cagatcctgt
gtcaggagtt gggattcttt gaaggtaagg gccttgacca 35640 ccgaattaag
gtaatcttgc tctgtggcag gccttgtttt cagtatttta agtacactgg 35700
ctcaggtaat cctcacaaca gccccaggag gaatgttcta ttacctccac tgtatagatg
35760 aggaacttga ggcacagaat ggttgccaag gtcacacagc tatattgggg
gttcataccc 35820 agccatccaa ctctgtctgt actctctgcc actctgcacc
cccagctcct gatccacttc 35880 ctgtttccat ccctcgattt ctgctgcact
caggggcccc tctccccctc ggcctgtgag 35940 atctgcttca gtaggctttt
ctccctgact cctccatccc tgtccttaca ggcagctgct 36000 tctctccggg
acacgagggg tccatacgga cactctctac tggctgggtt gcgcctaact 36060
cgtgattcct cctctgtttc agattcggag ccgggttgat gtcatcagac acgtggtaaa
36120 gaatggtctg ctctgggatg acttgtacat aggattccaa acccggcttc
agcgggatcc 36180 tgatatatac catcatctgt aagtccgaaa atgcctgtcg
tgtgtgcctt aggctgctgc 36240 ggaggaggcc agggctatat aagcagagtc
agtgactgac tgtgccctgc agtgttgatg 36300 gccatggaga ttccaccgtt
agagcttttt tctttgttaa ccttgaaggc aaatctggtt 36360 aggaagataa
ctttcaaaga gtcaccatct ggacattcat gcccatgtgc ttcaatcctg 36420
tatacaagca gtttagagta cagggaaggg aaggacatta tgaaagggag agggtgtgtt
36480 tggatccagc agctccatcc tcagaattta tctgaagaca ctgcaaaatt
actaagaatc 36540 actatgacaa gaatgaggat ggggtgatat ggcaaagttg
tgatcctgga agaccttcat 36600 ctcccatgtt gcccaactct gaacatgaat
ttggtgaact agttggttaa ggggatgatc 36660 ctccaagttt ctccctggtt
gagctccaaa aaccatgtaa gtttctcata gcaaaaccgt 36720 ataggtcctt
agggctttag ttggaatatt tgtgctgaaa tgctggaaag ccccatttgc 36780
catttttgta tttgcaaaat aatcatcaag aggggagaat gcattctttc atgaccactg
36840 accctctgaa aaggtcagga atttagtctg aagtaggcaa gcctcctacc
ccgcttctgc 36900 catgagcttg cacgcacagg cctgtcttga catttcttct
ttatagattt ctttttgaat 36960 atcttgaaat tgctttaaaa atatttaaag
aatgtagaat tatataaaat aaaaaggaaa 37020 taaccccaca cctcccacaa
aaccctgttt cctgcctttc tccacccact ctccagggta 37080 acacttggta
acagcatagt tgtatcaccc caggcctatt tttgagcata tcagcatttc 37140
aagaaatgta ttttttctca ataaaacatc ccttatagtt gaggagggga ggttatcatt
37200 cctgggtttt gttttttttt tttttttaat gtaatcctgg tacatcggta
atttgcattt 37260 tttattcatt aatatctttg gtatttctag tgttgggaca
cacaggtcaa cctcagtttt 37320 tgggtttttt tttttgtctt tttgtctttc
tagggccaca cctgcagcat atggacgttc 37380 ccaagctagg agtctaatca
gagctgtagc caccagccta cgtcatagcc atagcaacgt 37440 cagatccaag
ccgtgtctgt gacctacaag cacagctcat ggcaacaccg gatccttaac 37500
cactgaacga ggccagggga tcgaacacac atcctcatgg atcctagtca tgttcattaa
37560 ccactgagtc atgatgggaa ctccaacttc aactatttta atgtctgtaa
aacattccat 37620 ttggaaacca tttcatttgt aaagcaaaat gaaaacattt
tgttcatttt caacagagtt 37680 cgtagctgac ttctgttctg gaaaaaagga
aatggagcaa atttgagtga gaaagattca 37740 aagataactt ttcttttaaa
aaaaattata tcttggaaac ttctgggcta ttgattctga 37800 agactatttt
tctatatact gttttgatag caaagttcat aaatgtgaaa ggatcctgcg 37860
atgaatcttg ggaagcagtc atagcccaat atatctttgt tgcttttaaa atgagattta
37920 gtttactaaa tatttttctg atcataaaaa taacacagat ctaccgcaga
aaatttggaa 37980 aaaaaaaaac ttttaaattc aaaaaacagt taaaccacaa
atgatcccac catccagaga 38040 gcaatttgta ctttggtgtc tagttcatct
ttctttttct gtttacaagc acatatacca 38100 caagcatttt ttcaaaaaat
gaaaatggga taatactata catacgtctg tacacctgca 38160 tagttactga
acagtctttg atctaccctg taagtttcta acttttcatt atttgaaatg 38220
atgttttggc aaagaaatat gtaggtgtgt ctcgcacact ttcataatga tttcttagga
38280 taaatttctt aggataaatt cataatgatt tcttataata atccatactc
tgccaactga 38340 tcttcaggga agccaactcg ccttctcaga aataacatat
aacccattta cttgccctct 38400 caccaatact aggtcctaat gtttttgtgt
acagattcta tatttttaca tacaagaatt 38460 ccttaaagca aggcatgtca
cagaaaaata gaaggaagac acaattgtca tgtttaagga 38520 ctgcattctg
taccaaaaat gctaagttaa atgaacatct gaaacagtac agaaacgcta 38580
tctttcaggg aaagctgagt accaggtact gaacagattt tggcaaatac agcaggcatg
38640 gatgtttcca aaacatgttt ttctacttta tctcttacag gttttggaat
cattttcaaa 38700 taaaactccc cctcacacca cctgactgga agtccttcct
gatgtgctct gggtagagag 38760 gacctgagct gtcccaggta aagcatcctg
caggtctggg agacactctt attctccagc 38820 ccatcacact gtgtttggca
tcagaattaa gcaggcacta tgcctatcag aaaacctgac 38880 ttttggggga
atgaaagaag ctaacattac aagaatgtct gtgtttaaaa ataagtcaat 38940
aagggagttc ccatcgtggc tcagtggtaa cgaaccctac tagtatccat tgaggacaca
39000 ggttcaatat ctggcctcac tcagtcggct aaggatccag tgatgccgtg
agctgcagtg 39060 taggccacag acgtggctca gatctggtgc tgctgtggct
atggtgtagg ccggccccct 39120 gtaactccaa ttcgacccct aggctgggaa
cctaaaaaga ccccaaaaaa gtcgctttaa 39180 tgaatagtga atacatccag
cccaaagtcc acagactctt tggtctggtt gtggcaaaca 39240 tacagccagt
taacaaacaa gacaaaaatt atcctaggtg gtcagtgggg gttcagagct 39300
gaatcctgaa cactggaagg aaaacagcaa ccaaatccaa atactgtatg gttttgctta
39360 tatgtagaat ctaaattcaa agcaaatgag caaaccaatt gaaacagtta
tggaagacaa 39420 gcaggtggtt gtcagggggg agataagggg aggcaggaaa
gacctgggcg agggagatta 39480 agaggtacca actttcagtt gcaaaacaaa
tgagtcacca gtatgaaatg tgcaatgtgg 39540 gaaatacagg ccataacttt
ataatctctt tttttttttt gtcttttttg ccttttctaa 39600 ggctgctccc
gtggcatatg gaggttccca ggctaggagt ccaaacagag ctgtagctgc 39660
cagcctacac cagagccaca gcaacacggg aaccttaacc cgctgagcaa ggccagggat
39720 cgaacccgag tcctcacaga tgccagtagg gttcattaac cactgagcca
cgacaggaat 39780 tccagggtct gttgtgttct taaaacactt ccaggagagt
gagtggtatg tcataagtaa 39840 acaataaatg ttaaccacaa caagcttatg
aaataaacag gaaagccata tgacctacaa 39900 tcagtcattg ggagaatcca
caaaaggttg agcagaggat caattccagc tcacactcca 39960 gttttagatt
ctcccctgcc ttaaagcatc acagactaca taatctgagc tgaagaataa 40020
aaattaaaac tcaccccagt gcaaaacaga aatgaaaaag tattaaaacg aggttcatac
40080 tgttgttcat tagcaatatc ttttattcac aggggtgccc aacaacatga
aaaaatcaag 40140 aatttattgc tgctacgtca aagcttatac cagagattat
gccttataga cattagcaat 40200 ggataattat atgttgcact tgtgaaatgt
gcacatatcc tgtttatgaa tcaccacata 40260 gccagattat caatatttta
cttatttcgt aaaaaatcca caattttcca taacagaatc 40320 aacgtgtgca
ataggaacaa gattgctatg gaaaacgagg gtaacaggag gagatattaa 40380
tccaagcata gaagaaatag acaaatgagg ggccataagg ggaatatagg g 40431 47
1192 DNA Porcine 47 ctgccagcct aagccacagc cacagcaacg ctgggtctga
gccatgtctg cagcctatgc 60 cagagctccc cgcagcgccg gatgcttaac
ccactgagca aggccaggga ttgaaccctc 120 gtcctcatgg atagcagttg
agttgtttcc acggaactct taggggaact cctgattatt 180 ttttatttaa
atttatattt ctctgacttt ttcgtgtgct catcagccac tgactgtgta 240
tctccattag tcatggtttg ttaactctgt cattcaaacc ctcttcatcc ttgctacgca
300 gataacatca ttataataaa atcgtgcctg aagaccagtg acgcccccaa
gctaagttac 360 tgcttcccct ggggggaaaa agaagcaccg cgcgggcgct
gacacgaagt ccgggcagag 420 gaagacgggg cagaggaaga cgggggagca
gtgggagcag cgggcagggc gcgggaagca 480 ctggggatgt tccgcgttgg
caggagggtg ttgggcgagc tcccggtgat
gcagggggga 540 ggagcctttt ccgaagtagc gggacaagag ccacgggaag
gaactgttct gagttcccag 600 tcccgacgtc ctggcagcgc ccaggcactg
ttattggtgc ctcctgtgtc cacgcgcttc 660 ccggccaggc agccctggcg
gatcctattt tctgttcccc cgattctggt acctctccct 720 cccgccctcg
gtgcgcagcc gtcctcctgc agtgcctgct cctccagggg cgaaaccgat 780
cagggatcag gccacccgcc tcctgaacat ccctccttag ttcccacagg tgagaaggct
840 tcgccgctgc tgccgctggc gccggcagcg ccctccacgc acttcgtagt
gggcgcgcgc 900 cctcctgcat tgtttctaaa agattttttt ttatccgctt
atgctatcag ttactgagga 960 agtatttaca aatctactat tattttgaat
ttgccttttt ctccttatag tttatcagta 1020 tctcttgaga ctgttattgg
tgcctgcaaa tttaaaatga ttggggtttt atgaggaagt 1080 gaacctttta
tctttatgaa acgcctaact gaggcaatgt taattgctta aaatactttc 1140
tttattatca gtgtggccat gccagtgtcc tcttggttag aatttgcctg at 1192 48
39239 DNA Porcine misc_feature (15433)..(15433) n is a, c, g, or t
48 ctgccaaagc tgggagatgg gggaaagtag agtgggttat tgaaactgaa
tatagagttc 60 agcatctaaa agcgaggtag tagaggagga agctgtgtca
acggaaatac tgagctgggt 120 tcacatcctc tttctccaca cagtctaatg
ccttgtggaa gcaaatgagc cacagaagct 180 gaaggaaaaa ccaccattct
ttcttaatac ctggagagag gcaacgacag actatgagca 240 ggcaagtgag
agggggcttt agctgtcagg gaaggcggag ataaaccctt gatgggtagg 300
atggccattg aaaggagggg agaaatttgc cccagcaggt agccaccaag cttggggact
360 tggagggagg gctttcaaac gtattttcat aaaaaagacc tgtggagctg
tcaatgctca 420 gggattctct cttaaaatct aacagtatta atctgctaaa
acatttgcct tttcatagca 480 tcgaacaaac gacggagatc ctgttgtgcc
tctcacctgc cgaagctgcc aatctcaagg 540 aaggaatcaa ttttgttcga
aataagagca ctggcaagga ttacatctta tttaagaata 600 agagccgcct
gaaggcatgt aagaacatgt gcaagcacca aggaggcctc ttcattaaag 660
acattgagga tctaaatgga aggtactgag aatcctttgc tttctccctg gcgatccttt
720 ctcccaatta ggtttggcag gaaatgtgct cattgagaaa ttttaaatga
tccaatcaac 780 atgctatttc ccccagcaca tgcctaactt tttcttaagc
tcctttacgg cagctctctg 840 attttgattt atgaccttga cttaatttcc
catcctctct gaagaactat tgtttaaaat 900 gtattcctag ttgataaaca
gtgaaacttc taaggcacat gtgtgtgtgt gtgtgtgtgt 960 gtgtgtgttt
accagctttt atattcaaag actcaagcct cttttggatt tcctttcctg 1020
ctctctcaga agtgtgtgtg tgaggtgagt gcttgtccaa acactgccct agaacagaga
1080 gactttccct gatgaaaacc cgaaaaatgg cagagctcta gctgcacctg
gcctcaacag 1140 cggctcttct gatcatttct tggaagaacg agtgctggta
ccccttttcc ccagcccctt 1200 gattaaacct gcatatcgct tgcctcccca
tctcaggagc aattctagga gggagggtgg 1260 gctttctttt caggattgac
aaagctaccc agcttgcaaa ccagggggat ctgggggggg 1320 ggtttgcacc
tgatgctccc ccactgataa tgaatgaggg attgacccca tcttttcaag 1380
ctttgcttca gcctaacttg actctcgtag tgtttcagcc gtttccatat taggcttgtc
1440 ttccaccgtg tcgtgtcgtc aatcttattt ctcaggtcat ctgtgggcag
tttagtgcga 1500 atggactcag aggtaactgg tagctgtcca agagctccct
gctctaactg tatagaagat 1560 caccacccaa gtctggaatc ttcttacact
ggcccacaga cttgcatcac tgcatactta 1620 gcttcagggc ccagctccca
ggttaagtgc tgtcatacct gtagcttgct tggctctgca 1680 gatagggttg
ctagattagg caaatagagg gtgcccagtc aaatttgcat ttcagataaa 1740
caacgaatat atttttagtt agatatgttt caggcactgc atgggacata cttttggtag
1800 gcagcctact ctggaagaac ctcttggttg tttgctgaca gactgctttt
gagtcccttg 1860 catcttctgg gtggtttcaa gttagggaga cctcagccat
aggttgttct gtcaccaaga 1920 agcttctgca agcacgtgca ggccttgagg
tcttccgact tgtggcccgg ggactctgct 1980 ttttctctgt ccttttttct
ccttagtggg ccatgtcctg tggtgttgtc ttagccagtt 2040 gtttaaggga
gtgttgcagc tttatgatta agagcatggt ctttccttgc aaactgcttg 2100
gtttagaagc ctggctccac cacttagcgg ctctgtgacc tcggacacat ttcttagcct
2160 ttctgggcct cgctcttctt cctcataaag tgaaaatgaa agtagacaaa
gccttctctg 2220 tctggctact gagaggatgg agtgatttca tacacataaa
gcacttaaaa taatgtctgg 2280 catatgatac atgctcaata aatgtcactt
acatttgcta ttattattac tctgccatga 2340 tcttgtgtag cttaagaaca
gaggtcttta caggaattca ggctgttctt gaatctggct 2400 tgctcagctt
aatatggtaa ttgctttgcc acagactggt cttcctctcc ttcacccaaa 2460
gccttagggg gtgaacgatc ccagtttcaa cctattctgt tggcaggcta acatggagat
2520 ggcaccatct tagctctgct gcaggtgggg agccagattc acccagcttt
gctcccagat 2580 acagctcccc aagcatttat atgctgaaac tccatcccaa
gagcagtcta catggtacac 2640 tcccccatcc atctctccaa atttggctgc
ttctacttag gctctctgtg cagcaattca 2700 cctgaaatat ctcttccacg
atacagtcaa gggcagtgac ctacctgttc caccttccct 2760 tcctcagcca
tttttcttct ttgtacataa tcaagatcag gaactctcat aagctgtggt 2820
cctcattttg tcaatctaat ttcacagcct cttggcacat gaagctgtcc tctctctcct
2880 ttctgcctac tgcccatgag cagttgtgac actgccacat ttctccttta
acgacccagc 2940 ctgctgaata gctgcatttg gaatgttttc aatttttgtt
aatttattta tttcatcttt 3000 tttttttttt tttttttttt ttttttttag
ggccgcaccc atgggatatg gaggttccca 3060 ggctagggat ccaatgggag
ctgtagctgc tggcctacac cacagccaca gcaatgcaca 3120 attcgagcca
atctttgacc tacaccagag ctcacggcaa cactggattc ttaacccact 3180
gattgaggcc agggatcaaa ctctcgtcct catagatacg agtcagattc gttaacctct
3240 gagccatgat agttgttagt tactcattga tgagaaagga agtgtcacaa
aatatcctcc 3300 ataagtcgaa gtttgaatat gttttctgcc ttgttactag
aaaagagcat taaaaattct 3360 tgattggaat gaagcttgga aaaaatcagc
atagtttact gatatataag tgaaaataga 3420 ccttgttagt ttaaaccatc
tgatatttct ggtggaagac atatttgtct gtaaaaaaaa 3480 aaaatcttga
acctgtttaa aaaaaaaact tgactggaaa cactaccaaa atatgggagt 3540
tcctactggg acacagcaga aatgaatcta actagtatcc atgaggacac aggtttgatg
3600 cctggcctcg ctaagtgggt taaggatatg gtgttgctgc agctccaatt
caacccctat 3660 cctgggaacc cccatatgcc accctaaaaa gcaaaaagaa
aggtgctgcc ctaaaaagca 3720 aaaagaaaga aagaaagaca gccagacaga
ctaccaaata tggagaggaa atggaacttt 3780 taggccctat ctccaactat
cacatcccta tcaccgtctg gtaagaaatg gaaaaaatat 3840 tactaagcct
cctttgttgc tacaattaat ctgattctca ttctgaagca gtgttgccag 3900
agttaacaaa taaaaatgca aagctgggta gttaaatttg aattacagat aaacaaattt
3960 tcagtatatg ttcaatatcg tgtaagacgt tttaaaataa ttttttattt
atctgaaatt 4020 tatatttttc ctgtatttta tctggcaacc atgatcagaa
atctttaaac aatcaggaag 4080 tcttttttct tagacaaatg aaaatttgag
ttgatcttag gtttagtaca ctatactagg 4140 ggccaagggt tatagtgtga
ctattaaatc acagataatc tttattacta cattatttcc 4200 ttatactggc
cccacttgga tcttacccag cttagctttt gtatgagagt catccttaaa 4260
gatgacttta ttctttaaaa aaaaaaacaa attttaaggg ctgcacccat agcatataga
4320 agttcctagg ctagcggtca aattagagct gcagctgcca gcctatgcca
cagccacagc 4380 aatgccagat ctgagctgca tctgtgacct acactgcagc
ttgcagcaat gctggatcct 4440 taacccattg aacaatgcca gggattgaac
acacatcctc atggatactg ctcaggttcc 4500 taacctgctg agccacagtt
ggaactccaa agcagacttt attctgatgg ctctgctgat 4560 ctctaacacg
ttattttgtg ccatggtgtt tatcttcact ttactcaagt cagggaaaca 4620
cgaagagtct catacaggat aaacccaagg agaaatgtgc aaagtcacat acaaatcaaa
4680 ctgacaaaaa tcaaatacaa ggaaaaaata tcttcacttt caaaatcacc
tactgatgat 4740 gagtttatat ttccttggat atttgaatat tagctatttt
tttcctttca tgagttttgt 4800 gttcaaccaa ctacagtcgt ttactttgat
cacagaataa tgcatttaag ccttaaatag 4860 attaatattt attttcacca
tttcataaac ctaagtacaa tttccatcca ggtctgttaa 4920 atgcacaaaa
cacaactgga agttagatgt aagcagcatg aagtatatca atcctcctgg 4980
aagcttctgt caagacgaac tgggtaaata ccatcaatac tgatcaatgt tttctgctgt
5040 tactgtcatt ggggtccctc ttgtcaactt gtttccaatc tcattagaag
ccttggatgc 5100 attctgattt taaactgagg tattttaaaa gtaaccatca
ctgaaaattc taggcaagtt 5160 ttctctaaaa aatcccttca ttcattcatt
tgttcagtaa gtatttgatg agaccttacc 5220 atgtgtaaac attgcactag
gtattaagaa atacaaagat ggataagata gagtcggcgt 5280 aaatgagatg
atataatgag acgttataat gaaactcaca attccagttg ggaaataaag 5340
tccttcaaat tccatgactc tttctggcac acgttagagg ctacagcttc tgtgtgattc
5400 tcatgctggc tccacttcca ctttttcctt cttcctactc aagaaagcct
atagaaatat 5460 gagtaagaag ggcttaatca taggaataaa tttgtctctg
ttctaagtga ttaaaaatgt 5520 ctttatcagt ataaaaagtt acttgggaag
attcttaaaa ctgcttttac acactgttct 5580 agaatgactg ttatataaat
aaaaaagtag atttgatcta acacaattaa atgacctttg 5640 gaaatattga
ctaattctca ccttgcccct caaagggatg cctgaaccat ttccttcttt 5700
tgccagaaag cccccaccct ttgtctgttg acctagccta ggaaatcttc agatcacgtt
5760 gttagcacga actggttaca tgtgctgtac aaatactatt taattcatct
gattaaaaaa 5820 aaagagataa gaagcaaaag tttgactatc ttaaactgtt
tgcgtaggtg agaggacaat 5880 tgaccatcta ctttatgagt atgtaaccca
gaaacttaaa gctccttaag ggagctaagt 5940 cttttggata agacctatag
tgagaccttt tagcaaaatg gttaagactg aatggagctc 6000 actagcgtgg
gttcatatcc tgatgctcaa acacgcaatt aaatgacttt aggtgggtta 6060
gtctctgttc cttagtttcc tcaatgggag ataatattgg tagtagcgat tttactgggt
6120 tgttgaaaga acatctgtta aatgttcaga acgtgttacg acagagtaca
gagtaatgat 6180 ttgcttgtat atgtatgact caaatagtct gccatatgcc
ttgtgactgg gtcctgtgga 6240 gcaggaagga gggatttccc acccagcaga
aagttgggta aactggaaaa tagactgagg 6300 ccaggaaatg atgcaaagcg
ttgatgttca ctgccacggc aggtgaaggg cagggccaga 6360 gttgtcagta
gggtcagggg aggactggaa ataaccaaga cccactgcac ttttcagcct 6420
ttgctccagt aaggtaatgt tgtgagagta gaaaattttg ttaacagaac ccacttttca
6480 gtacagtgct accaatactg tagtgatttc ataccacatc ccaagaaaga
aaaagatggc 6540 tcaatcccat gtgagctgag attatttggt tttattgtta
aataaatagc attgtgtggt 6600 catcattaaa aaaggtagat gttaggaaag
tagaaggaag aagactctca cctacatttt 6660 catcactgtt ttggtatctg
ccagttgtca ccttggtccc cttccccgcc tctcccctgc 6720 ctcctcttcc
tccttctcct ttttttggaa tacaattcag gtaccataaa atttaccctt 6780
ttagagtgtt tgactcaatg gtttttagta ttttcacatg ttgtgctatt actatcacta
6840 tataattcca ggtcattcac atcacccccc aaagaaacct tctaactatt
agcagtccat 6900 tcccttcttc cctcagcccc tggcaaccac taatctactt
actgtctcca tggatgttcc 6960 tatattgaat caagctagca taaaccccac
ttgctcatgg tcataattct tttttatagt 7020 gctaaattac atttgctaat
attcaattaa ggatttctat gtccatattc ataaggaata 7080 ttggtgtgta
gttttctctt tgtgtgatat ctttgtctgg ttgggggatc agagtaataa 7140
ttactgctct catagaatga attgagaagt gttccctcct tttctattta ttggaagagt
7200 ttgtgaagta tattggtatt gattcttctt taaacatttg gtcagattca
ccagtgaagc 7260 catctgggcc atggctaatc tttgtgaaaa gttttttgat
tactaattaa atctctttaa 7320 tttgttatgg gtctgctcct cagacgttct
agttcttctt gagtcagttt tgttcatttg 7380 tttcttccta ggactttctc
cctttcattt ggattattta gattgatagt aatatccccc 7440 ttttaattcc
tggctgtagt aatttgggtc ttttctcttt tttcttggtc agtttagcta 7500
aaggtttgta attgtattaa tcttttcaaa taactaactt ttttgttttg tttgtttttt
7560 gttttttgtt ttttgttttt tgtttttttt tgctttttaa ggctgcacct
gaggcatatg 7620 gaagttctca ggctagaggt ctaatcggag ctacagctgc
tggcctatac cacaaccata 7680 gcaatgccag attcaagctg catctgcgac
ctacaccaca actcggccag ggatcacacc 7740 cgcaacctca tggttcctag
tcggatttgt taaccactgt gccacgacgg gaactcccgc 7800 ccattttttt
taacacctca tactttaaca taaagatggg cttcacatgg actgatagct 7860
caaatgagga aggtaagact atgaaagtaa tggaagaaat gtagactatt tttgtgacct
7920 agagattact gatacttctt gacttttcaa acaatacttc aaaagtacag
cccaaaggga 7980 aaaaagaaag aaaaaagaaa cacacatata cacaaaccta
gtgaataaga tatcatcgat 8040 acactacaga tttctatgaa ctggaagacc
ccatggacaa agttaaagaa catatgatag 8100 tttgagtgat tattttgcaa
tatttacaac caatgaggga atattatcca gcttatagga 8160 ggaagtaatg
caaatcgaca agaaaaagat aggaaaccca atataaaaat taagaaaata 8220
caaaaattaa gaaaggatat gaactagcat tttacaaaag aaaaatctcc aaaagtcaat
8280 cagcacatga aaatatgctc aaacctatta attattagaa aactacagac
tgaagcaatg 8340 aggtgcttta ctttacatct ttttgactga taaaaagtta
gaaacaaagg tgatatcaaa 8400 tgtcagggat aaaaggatat agaaatcgtc
atgcctgtgg tgggagtatg gccggtgcag 8460 tcatgtggga aggtaatctg
acagtggtta ggcagagcag gtttatgaat acactgtggc 8520 ccatcaatcc
cacgcctgtt tatgtaccaa agaaatcctg ttgtggcaga atctatgggt 8580
ccacccctgg gagcatgaat taataaaatg tggcaccagg gtgtgtgaaa ctccagctag
8640 agatgagatg tccacatggc aacatgaatg catcttagaa acatagattt
gagtgaaaaa 8700 gagtaagaaa cagccgggaa acccaatacc atttataaaa
attaaagatg cacacataca 8760 atgtagtaaa tattttgcat gaactttcaa
atggttgcct acaggggggg agagtaaaga 8820 agagtagaaa acaaagataa
agggagtaag taagtagctc tgcctggact gaatataatg 8880 tgtcatgaac
tgagaaatat ggttaacata atcctcttaa cttgaggtcc taaatgaatg 8940
aatgagtcca ctattcattt acccattctt taatgtgtat tgcattataa tccatttttt
9000 tagaaccaac gaattttgtt cccataacta ctaatcagcc tgccttttct
ccctcattcc 9060 cttatcagct caggggcatt cctagttttt caaacgttcc
tcatttgaac caaaaatagc 9120 atcattgttt aaattatact tgttttcaaa
tacgatgctt atatattcca agtgtgtttg 9180 cccattttct taggtggtag
aaatttttca ttctactttt ctatctactc agattttccc 9240 gttggaatta
tttccattgc tattaaactt agaagtcccc cctgtgatat gccatttttt 9300
tcatactttt taagcacttg gttgcttttc tttgtgtctt taagcaccta gaatacttat
9360 aaccattgca cagcactgtg tatcaggcag cccttcctct tccactaatt
tatggtcctt 9420 ctcttagact atattaaact gttatttaat taggatcctc
tcttcgtcct tatgatttaa 9480 ttattatagt tttctaatat gtttttatta
taattcctct tcattattcc tccctattaa 9540 aaattttaat gaattccatt
tgtttgttct tctagttaaa tattaagtca taatccaaat 9600 aacttagatg
tcattagttt atgtggtcaa agtaaggata ccacatcttt atagatgcag 9660
gcagttggca gatgtcatga ttttcttcag tgcataaatg caatttatct ttgagcaagg
9720 ggcataaaaa cttttatggt attggctttg aaataatagt taagaactgc
agactcagtt 9780 tttcctgctt ttcttgaaaa agaacacttc taaagaagga
aaatccttaa gcatggatat 9840 cgatgtaatt ttctgaaagt ctcctgtaat
tccttgggat ttttgttgtt gtttgttggt 9900 cggttttttt gggtttttgt
ttgtttgttt tgttttgttt tgttttgctt ttagggctgc 9960 acctgtggca
tatggaagtt cccaggctag gggtccaact ggagctacag ctgccagcct 10020
actccacagc cacagcaaca tgggatccta gctgcatctg tgacctaacc acagctcttg
10080 gtaatgccag attgttaacc cactgagcaa tgccagagat cgaatctgcc
tcctcatgga 10140 cactagtcag attagtttct gctgagccac aatgggaatt
cccaattcct tgtatttttg 10200 aactggttat gtgctagcat ataattttgt
ttcttgaatc tttgtgggtt tttttttttt 10260 tttttttttg tctcttgtct
ttttaaggct gcacccacag catatggagg ttcccaggct 10320 agaggtcaaa
ttggagctac agctgccagc ctacacaaca actgcagcaa agtggggccc 10380
aacttatatg acagttcgtg gcaatgccgg attcctaacc cactgagcag ggccagggat
10440 cgaacctgag tttccagtca gtttcgttaa ccactgagcc atgatagtaa
ctcctgtttg 10500 ttcagtcttg aacctccttt ttaattcttt attccttgag
ggtgaaataa ttgccataat 10560 aatactatca tttattacat gccttctctg
tgctaggcat agtgacactt taggatttat 10620 tatatcactt aatccctaca
acaactctgc aaagtatgta tcataatcct atttgacaga 10680 tcaggaaatt
gcagcccagg atgcagataa tatgcatcca tcacaagtga ctagatatag 10740
tccctctgct attcagcagg gtctcattgc ctttccattc caaatgcaat agtttgcatc
10800 tattgtatat gtgttttggg gtttttttgt cttttttttt ttttttgtct
tttctggggc 10860 ctcacccttg gcataggtag gttcccaggc taggggtcaa
attgaagctg cagctgccag 10920 cctacaccac agccacagca actcgggatc
tgagcctcat ctgcaaccta caccaaagct 10980 cacggcaaca ccggatcctt
aacccactga gtgaggccag agatcaaacc ggcaacctca 11040 tggttcctag
tcggattcat taaccactga gccacgatgg gaactcccta aatgcaatag 11100
tttgctctat taaccccaaa ctcccagtcc atcccactcc ctcctcctcc ctcttggcaa
11160 ccacaagtct gttctccatg tccatgattt tcttttctgg ggaaagtttc
atttgtgcca 11220 tttttcattt tacgggtaat ttttacttca gtttcttcca
ctagcagttg tcttaaagtg 11280 agtataatta atattcattt ggaaaatgta
agcaaaacat tttttaaagg gccatgccca 11340 cagcatatga aagtttctgg
gccaggggtt gaatccaggc tccaagttgc agctgtgccc 11400 tacactgcag
ctgggcaatg ctggatcctt taacccactg tgcccggcta gggatcaaac 11460
ctgcatttcc acagctaccc gagccattgc agttggattc ttaacccact gcactacagt
11520 gggaactccc acaaaacatt ttttaatgtc ctttgaataa agtaggaaag
tgctcgtctt 11580 tgagggcagg gcggcaatgc catttccaca aggtttgctt
tggcttggga cctcatctgc 11640 tgtcatttag taatgaataa aattgctgac
agtaatagga ttaactgtgt gtggagatag 11700 ccagggttag agataaaaac
actggagaag tcaaataagt tgctcgaggt cctctagcta 11760 ataagctatt
aagtgggaga gtgagggcta gaaacaggcc atctgtctcc caagcacatg 11820
tccattagtg gtttgctgat agccttccag aacaacagag aggactctca aacatggtct
11880 tgcctccctc caattgatcc cctccatgtg cctcacagcg ggtctttcta
aaattaagtt 11940 ctgattttaa ttctcccttg ctatagcact taggtatggc
tttcagccgt gcaataaaaa 12000 gcaggcaaga gtggctcaat catataggag
gttgtttttc ttagatccca agcaggtaat 12060 cctgggcatt atggttgttc
tgcgtttatc aaggagccaa attctctatc acctcctgtt 12120 ctatcctcct
cagtatctgg ctctattctt cagcatctca agatggcttg tgctcctcca 12180
agcatggcag tcaaattcca cacaagaggg ggaaatatga agggcagaca gtgctggtct
12240 cctgagctgt ccctctttgt cggggaaata aatgtattcc ttcaagtccc
gtgagacttc 12300 tgaagtagac gtctgcttac gtctcaccca ccagaactat
gtaaactgca catagtgcta 12360 ggtctacata gccactcata actgccaggg
ggtgggaaat ctttaaatag gtgtaccacc 12420 acacaattag gatgctaata
gtaagggaga aggagagaat aggttttgcg caagccacca 12480 gcatgcctgc
cacaattgct taaaattctt cattgacccc tcattgccac aggatgaaat 12540
ccaaacgcct tcttagttgg gaatctgacc tacctgtctc tcccacctgg ttcagacacc
12600 attctccttg gtcataaaat tccagtcatt tgtgaacatc cagctccccc
atgcctccat 12660 gcctttgcac atgctgttct tttatctttt atgttgtcct
tttatctttt atccaaaaga 12720 gatatcccat catcacatct cttttgtcag
cccccaaata ctttgtcttt caagttcagc 12780 tggaggatta cctcctattt
gaaatcagct ttgtctctta caaccaaaca aggttttcct 12840 tccgagacac
tcccacagca ccttgaactc atctctatca atcattcatt tgattgtaat 12900
gaagttgttg gtggtatgcc tgtgtctctg acacatctgc gatctcatga gttccttaag
12960 tggaatgtga atagcgggat gaacagtatt ggtcttcagc cctcatctct
gcagatgttg 13020 cttgacccaa atgagcgttg ccttttattt tgattttgct
ttgatttgtc tactccatgt 13080 acttgagcca tgcatttctg tcttagcgat
gctttttaaa agtcattttt tggttgatta 13140 tccagatttg tccacctttg
cttctagttg tagaaaagga tgaagaaaat ggagttttgc 13200 ttctagaact
aaatcctcct aacccgtggg attcagaacc cagatctcct gaagatttgg 13260
catttgggga agtgcaggta aggaaatgtt aaattgcaat attcttaaaa acacaaataa
13320 agctaacata tcaatttata tatatatata tatatatatt tttttttttt
tttacatctt 13380 atattacctt gagtattctt ggaagtggct agttaggaca
tataataaag ttattctgaa 13440 gtcttttttt ttctttttcc atggtgagca
gtggcttgat gtggatctca gctcccagac 13500 gaggcactga acctgagccg
cagtggtgaa agcaccaagt tctagccact agaccaccag 13560 ggaactccct
attctaaatt cttgagcaca ttatttagga acctcaggaa cttggcagga 13620
ttacaggaaa tatatctaga tttaaaaaaa aatcttttaa cagaggtccc aaaggagagt
13680 catgcacagc tatgggagga agttcagaaa ctgcccttgc taccagatca
ctgtcagata 13740 aaatggccag ctacatgttt ctgcacattg ccctaagatc
tttacaaact tttctgtgca 13800 tttttccact tttaaaagaa aatttcgggg
ttcctgttgt tgctcagtgg ttaacgaacc 13860 caactagtat ccatggggac
aggggttcga gccctggcct cactcagtgg gttaagaatc 13920 tggcattgct
gtggctgtgg cgtaggctgg cggctacagc tcagattgga cccctagcct 13980
gagaacctcc atatgccgca ggtatggccc taaaaaaaaa aaaaaagaga gagagagaat
14040 ttcctccaga aaaaacactt tggtagtttg ggagaagtaa acaaccaaaa
attaattttt 14100 ctggagtatt cgggaagctt gtaaaaatgg gctcttactt
ttttgaggag acaaatggga 14160 acctacccag aagaggcaca atcacctgca
tttgatttct tgacctctcc ctaccttctt 14220 tgctggcttt ccacatttgg
atttctgtga ccttatctct gctccttggt gttttcattt 14280 ttcctgtgga
cgtgccagac tatgggaagg gagtaaggcg ttgatttaga
atcctgtagt 14340 ctctgcctgt ctctagtcat tgttttcacc cttctcaaag
gaccttgaca tcctgagtga 14400 gtccgcaagt aatttagggg agaagcctta
gaagccagtg cagccaggct acatgactgt 14460 gtccacccac tggaaccagt
catttttata cctattcaca gcccccctac catttaaatc 14520 cccagaggtc
tgccataaca tctgtaactc cctttcctgg taaattgtgt tctaaaagac 14580
tggtaacaaa agatattctg tggtacagag cataattaaa tacctgggag ctgatttgag
14640 tggggtaaat caactggttt gacccctaaa acccaccatg agcatttctg
ttctaataaa 14700 gtaatgcccg tgctgggaat tgtgttctac ggaaatgctc
ctgctgtgtc tttcttgagt 14760 cctgtgtcat tgaacatgct taggagcaaa
ggtcccccat gtggcttgtc tgctaaccag 14820 cccagttcct tgttctggct
ggtaatgatc cgatcatctg aatctcactg tcttccaaca 14880 gatcacgtac
cttactcacg cctgcatgga cctcaagctg ggggacaaga gaatggtgtt 14940
cgacccttgg ttaatcggtc ctgcttttgc gcgaggatgg tggttactac acgagcctcc
15000 atctgattgg ctggagaggc tgagccgcgc agacttaatt tacatcagtc
acatgcactc 15060 agaccacctg aggtaaggaa gggtgagccc tcaactccga
agaaaatgct gcaataaaag 15120 cactgttggt tttcagcttt ttttgtaatc
actgctcatt ctgaggtaga ttcgcttggg 15180 ctgataaaaa gagaactaat
tcagataaat gcttgcattt gcatagcctc tttttttaaa 15240 aacttttttt
tttttttttt ttttttggct tttcagggct gaacctgtgg catatggagg 15300
ttcccaggct aggggtcgaa tcagagctgt agccccgggc ctatgccact gccatagcaa
15360 catgcatagc ctccttttta aagtgccttc ctgttttata ccattgggat
gtgagaagag 15420 ctattgtgga aangagcatg gggtnataac cctggacctc
tcacgtccta ccctcaggnt 15480 agtgggaaaa ctctgagttt aaggacatca
aagtgactcc tttttagtta cattatggng 15540 gaatcagcnc atatttttac
aaggggcgga gngtaanctg ttggagttta caagacatat 15600 ggtggcattg
caactactta accctactat tatagcacaa aagcagccat agtcggtcct 15660
gaaggagcct gatgccttca gctttatagg caatgacgtg tgaatatcac aaacagtttc
15720 ctgtgtcacc aaacatgatt gccttttgat ttccctttca accctttaaa
aaaaggtaaa 15780 agcccttctt agcattcagc agcaggtcgc tgtgttttgc
caactcctga tctgtagcat 15840 ttcgacaaca ctgagctctc aacttttgaa
ccctgagtcc accacatcct tcagtgaaac 15900 cagagccatg tgatactaag
gatagaaacg gaaacttcct gaatccaggc gatcaaatag 15960 gagggagaaa
gaggaacttt cattgacaaa accacaaata ttgtgaatgg actgttacaa 16020
atattgtgaa tgctcctatt cccaaccccc tggcttcatt acagggtcct atgtgttcat
16080 ccttattgag aaatttgtat tgctactgcc aggttgccaa tacccagcgg
tgcccatggt 16140 gttctaaaat gaagcaattt caactttatt tttttttcct
gtgactttac atgacaagtt 16200 cacatgaagg atatactttg atagtaatgt
ccatggttag ggaatataca ttgtttgctg 16260 gttgactggc ccctggattt
ttctattgaa agtccatgag atctcgaagg cacaggtgtg 16320 ttctctcgct
ttttaaggaa agggtttaaa aacttaagta attaacagct ttagtaacaa 16380
attacctata acacacttaa aaaccgaata ccacccactg gagtattgtg ctacgattaa
16440 aaatctactt gtctactaca tgatatcttt gtcccacaga aggttctgga
accaaacttg 16500 taatttcagg attatgagag ccctgagttc acgcattgtg
taataactat gttgtgtggt 16560 agtcaatttg tacagcttgc ttagagagaa
caatgtcaag ttaaggaggc gattgcttta 16620 tagtgcctgt cacaagatgc
cattgccatt gtcctagcaa gagatattct atgggagtat 16680 actacatttt
agtgaggata agaacttttt atggcattta gtccggtcat ttcccaacca 16740
ctgtcctgaa aaccaatttc attttgattt caggggcttg tgtgggcaaa gttgccaggc
16800 attaaaaagc cacttctcaa ctgtagtatc acaatgcttt agttgggtag
tgtattgcag 16860 atagcttatg gctgaaaagt taccaagcct tgcagttttc
actcctttga gtttatttcc 16920 ttgacagaat tgaccctgag ttttttgact
cttacctgct caactaataa acaccagagt 16980 catttatctc cattgctctt
gtctgacctt tatttaccga ataatgcctt atgggttcac 17040 aaaaacaagg
ggggaggggg ccagcatgcc ttagaaactg tctttagtca agaaatgnga 17100
ttttattatg taaatatatg agtattataa tagatagtgt tattaataga caccagcaag
17160 aattgtcaat aatttaaaaa tcacaaatta aaatacatcc atgttagnat
catttatcct 17220 aactcccaaa gccctttaaa gtggaagatt tagatgttaa
cccagagatt aaagacatgt 17280 tcaaagaatc cttgattttt ttttgaatcc
cttgttttta gagaagaaaa cctaatgatt 17340 ttccccctct ggattctaca
tattaaatat agttttggaa cttgaatatt agtatggtta 17400 ataagtgctg
atatgctgat tttgtttata tttttcttat gagtaaatat cctatatcac 17460
cagacattat agtctatgta caaatatgat tcttaaacct gatagcacat tcattagagt
17520 tggaattgcc tttttttttt ttttttttac agttgcacct gcaacatatg
aaagttccca 17580 ggctaggggt tgaatccaag ctgcagctgc caccctacat
tacagccgta gtaacagcag 17640 atccgagctg catctgcaac ctatgctgca
gctcagggca atgccagatc cactgagtga 17700 agccagggat ggaacttgca
tcctcataga gacaacgtcg tgtccttaac ccactgagcc 17760 agaacaggaa
ctccagaatt tcctttcaat agaagaagca ccaagtttag gatcagaaag 17820
cctgaatttg aataccaatt tactatttgt tagtcatata tttctgagtg tgtttcctca
17880 tttattaaaa gcagactaaa agatgagagg gtcttttgtt gagaatcaaa
tacaataaca 17940 tgtgaaagtg tgtaacacta tgattgaaat atacctacac
agccatttat ttgtttattg 18000 ttcatgtttt gccacccaca cagtagtata
taatcctttt atgtaataaa tgctaataat 18060 gaaagttggc aacttatgta
agtactcaaa atgctggagg tcatgggata ctgactggga 18120 tactacagag
gtaatgtcat ttcctctgcg ctaaacttat tgtcttgtag ttagggactg 18180
actctcttta ggacaaggag ttcattctgt ataccatgtg tggctatcac ccttcgaagt
18240 tgaaaaactg ccccagggtg ggcacccatc cgttctctta gatatatggc
cgagaccttt 18300 ctctcactgg gagggaacca cactgaggaa tgagaaaaaa
aaaaggaaaa tcaagatgaa 18360 accagaaacc tctttggcat aacttctcca
ctctgtactt tttgttagaa ctacccttgc 18420 acaaagcagc atcagtgtgg
aagacagaat ttgcacacct ggtttgatat acatgccgtg 18480 gtatatggga
tgttctaaca ataaagagga ctctcccagg aaatctcctc actgttatag 18540
tcagccttga ggaaagagct cttcttttgg actctgggga gagtctagtt tttcagttcc
18600 ttgcttctcg gtcaacgtgt tggtgtaagg atcacactct ctcttatact
agataattct 18660 attttttcac ctttcaacct gtctatcctt ctgaccctag
ttacccaaca ctgaagaagc 18720 ttgctgagag aagaccagat gttcccattt
atgttggcaa cacggaaaga cctgtatttt 18780 ggaatctgaa tcagagtggc
gtccagttga ctaatatcaa tgtagtgcca tttggaatat 18840 ggcagcaggt
ctgtgttctt tccacatgtt tgggttatcc tttctgggat aaatttgagg 18900
cgagatagaa actttaagac taaagaaaca atggcctact ttttttgtac atggtcctgt
18960 gtaaatctct atttgagctg aaataagatg gtcttcctct ccaattatcc
atggtatgac 19020 tctgatggat aacaaatcca gttctgaaaa aaggggattt
ctttccagaa gagaggacag 19080 tttcttcaaa tattgaatta aaagcaaaat
agatgtaaac cgttgttggt tttattgttg 19140 aattccaggt agacaaaaat
cttcgattca tgatcttgat ggatggcgtt catcctgaga 19200 tggacacttg
cattattgtg gaatacaaag gtattttctt gccctcatca gcatgaaatt 19260
gctcttggta gaaaggataa taatagttat ccaaaacatc atcctatgtt catctgtttc
19320 ttccctcttc attttccata gagtacagta tattctatct ctgtcttagg
aaaatggact 19380 gtcattcata taatcttaca gagaatcaat tagtaatgta
ctctatgccg tgacaggtgc 19440 gaaggttttt tttgaaggca acagataaaa
atatcctata tttcacctat tgtaatttcc 19500 ttaaaactga cattattgaa
taaatgtttt actttcatct tgaatattat tatgttatgg 19560 aatcatacac
tttaccccaa taatcatcga aaagaatttc caaaaggttg agagagttgt 19620
gttgatctga ttactttcct ctgcatcctt tgagcttaac ctttgaatat agtttgctaa
19680 ggaaagtagt ctgtttatga tcctggagtg gaatcaggct aagtgtcctc
attcagaacc 19740 cactgaatca gacagaatga atttatttcc ttgaaagttc
aaaatgtgtc actcaagagt 19800 ataaattttc aaatcttact ctctcttttc
cttggatgtg agcaattctt cgataattga 19860 atgaggcaga ttatatagac
ttacatggaa gactgttggc ctgagaattc aaactatggt 19920 gttcaagact
tcacngngag tccgatgcca tttgtttccc acaggtcata aaatactcaa 19980
tacagtggat tgcaccagac ccaatggagg aaggctgcct atgaaggttg cattaatgat
20040 gagtgatttt gctggaggag cttcaggctt tccaatgact ttcagtggtg
gaaaatttac 20100 tggtaattct ttatatcaaa atgatgccaa ggagttggca
tggcactttg ctaaatgctg 20160 tgtgaatcaa tacaaagata attaggacat
ggttcttcct cacaagaggt gtgcaatctt 20220 attgggaaat catacttgca
agtcacaaat atagactaaa gtttccagct gagaatatgc 20280 tgatggagca
tgaaacacta aggagacagg gagaatctca ggaaaaatca agaataattt 20340
ggatcaaatg gattcctgac atagaacata gagctgatca gaaagagtct gacattggta
20400 atccaggctt aagtgctctt tgtatgtggt tcagaacaga gtgtgggcag
cctgaggggg 20460 atacataccc ttgacctcgt ggaaagctca tacgggggag
ggatgaggct aaggaagccc 20520 ctctaaagtg tgggattacg agaggttggg
ggggtggtag ggaaaatagt ggtcaaagag 20580 tataaacttc cagttacaag
atgaataaat tctaggggta taataacagc atggcactat 20640 agatagcata
ttgtactata tactggaagt gctgagagta gatcttacat gttctaacca 20700
cacacacaca cacacacaca cacacaccac acacacacac cacacacaca cacgtgcaca
20760 caaacagaaa tggtaattat gtgaggtgat ggcggtgtta actaacttta
ttgtggtcat 20820 catttagcca tacatgcatg tcatgaaatc accatgttgt
acaccttaaa gttatgtaat 20880 actagatgtc agttatatct caaagctaga
aaaaatgtgg ggaccaaggc agaagctctt 20940 ctgctctgtg tctaagggtg
gttctggggc tgggatgggg aggatggtta agtggtatat 21000 ttttttcata
cctttgctca gtactatcat tgtaagtgtt caatatatgt ctgcttaata 21060
aattaatgtt tttagtaagt aatctctgtt tagtaatgtg tcagaaatgc cctacttgca
21120 ataggaagaa aacctgtcca gtcccttcct tttttctgta agtctgattt
cattgcctcc 21180 cagaatgcat caccatgtga gagatagagg gaaggtgctg
tccttatggg gttaacagtg 21240 tgactaggga ggcaaaatat acctactaaa
gggtggtagc ataattcagt tcttatgtga 21300 gtatgtgtat gtgtgtgagt
atgtgcacat gcacatacat tttaaaaggt ctgtaatata 21360 ctaacatgtt
catagtggtt acacctagct tataggtaac attttttccc ctgtatcctt 21420
gtttgtgttt atcaaatttt cataacagta atggtagaag gagtacctga catggtacca
21480 tacatgctng gncctgccta atttctcnat ttcctttatt gcccataccc
ccattgcttg 21540 acaagcataa gtccatactg gcttgttttc gttcctcaga
ctcagtacac catgtagctc 21600 catgccctgg gtctttgtat gtgctatttc
tactgcttag agtgctattg cccctgacca 21660 ccacgtggtc agcaacttct
cttctgcgtc tgtgtctatg gtctatgatt ccagatgtca 21720 tcttcactaa
ctacccttct aatatgccct tccatcccac ccgtcctcat ccttacccca 21780
gccactctct atttggtggc tctgttttat tttcttccta gctcatcact ctttgaaatg
21840 aacttattta cttattcaat atttgcttct ttcactagaa tgaatgctcc
atgagagcag 21900 ggacctgctt tatcttgctc gccactgtat tcacagtgcc
tagaactacg tctggcacat 21960 agtaggtgct caataaatat cgatcaaatg
aaagaatgag caaacgaaca aatgaacaac 22020 acgtgaggta ggcatcatga
ttccatcaac agaggagaaa accagactta aagnaatgaa 22080 gtggnggagc
tgcatttgat cttgactgac tccaacatcc atgctcttga ccactgtgca 22140
tctccagagt gtaatgaaca tactttactt ttatattcca ccaaaataac aaagccatgc
22200 ccatgttagt agagagttaa tcgacagtgc ccttaaaata tgcatgcacc
cagggtacaa 22260 ctatgcatgc tgccctgtgt tttcagttgg atccaaatga
attgccgtaa acaaagaggg 22320 gattcaatgt ctttgactag tttgggatat
tttcctagta accaactttg caaaataaag 22380 ccactaatga caaggagctt
tgttctactt ctgcatcact caactgtcaa tttttatctc 22440 ttgcaagact
tctaatctac tagaactttt gtttttctgt gatttctgaa cagagaagac 22500
taatccaaac cctgtcattc cagaggaatg gaaagcccaa ttcattaaaa cagaaaggaa
22560 gaaactcctg aactacaagg ctcggctggt gaaggaccta caacccagaa
tttactgccc 22620 ctttcctggg tatttcgtgg aatcccaccc agcagacaag
tatggctgga tattttatat 22680 aacgtgttta cgcataagtt aatatatgct
gaatgagtga tttagctgtg aaacaacatg 22740 aaatgagaaa gaatgattag
taggggtctg gagcttattt taacaagcag cctgaaaaca 22800 gagagtatga
ataaaaaaaa ttaaatacaa gagtgtgcta ttaccaatta tgtataatag 22860
tcttgtacat ctaacttcaa ttccaatcac tatatgctta tactaaaaaa cgaagtatag
22920 agtcaacctt ctttgactaa cagctcttcc ctagtcaggg acattagctc
aagtatagtc 22980 tttatttttc ctggggtaag aaaagaagga ttgggaagta
ggaatgcaaa gaaataaaaa 23040 ataattctgt cattgttcaa ataagaatgt
catctgaaaa taaactgcct tacatgggaa 23100 tgctcttatt tgtcaggtat
attaaggaaa caaacatcaa aaatgaccca aatgaactca 23160 acaatcttat
caagaagaat tctgaggtgg taacctggac cccaagacct ggagccactc 23220
ttgatctggg taggatgcta aaggacccaa cagacaggtt tgacttgaat atttacaggg
23280 aacaaaaatg atttctgaat tttttcatgt ttatgagaaa ataaagggca
tacctatggc 23340 ctcttggcag gtccctgttt gtaggaatat taagtttttc
ttgactagca tcctgagctt 23400 gtcatgcatt aagatctaca caccaccctt
taaagtggga gtcttactgt ataaaataaa 23460 ctattaaata agtatctttc
aactctgggg tgggggggga gactgagttt tttcacagtc 23520 ctatataata
attttcttat cctataaaat aattaggagt tcccgtagtg gctcagcaat 23580
agcaaacccg actagtatcg atgaggatgc gggttcgatt cctggccccc ctcagtgggt
23640 taaggatctg gcattgccgt gagctgtggt gtaggtggca gacacggctc
agatcccacg 23700 ttactgtggc tgtggcatag gccagcagct ccagctctga
ttagaccctt agcctgggaa 23760 cttccatatg ctgtgggtgt ggccttgaaa
aaaaataaat aaataagata attactcaaa 23820 tgttttcctt gtctcagaac
cttacttcag gataaagagt gagaaagttt tttttatgaa 23880 gggccattat
tacagctcaa aaataagttg tcttcagcaa gtagaaagca ataagcctga 23940
gagttagtgt tcctatcagt gtaaatatta cctcctcgcc aatccccaga cagtccattt
24000 gaacaattaa cggtgccctg ggagtacagt tcagaaacat taatgtggat
gttccagacc 24060 tgtattttta taagtacttg tcttgagccg gatggaacca
tcattcctca ccattattta 24120 gaagtggact gtgactctgt tggagatcag
ggcacacggt taccaaaagc acacccttct 24180 cctggcctta cctttgcaaa
gctggggtct gggacacagt cagctgatta taccctttta 24240 ctaacttccc
acagctcaaa tctggtcaat tctccttcac aaatctctta aaaatccatc 24300
actcacctcc agcctcttct gctgtggcct tgattcagcc tctcacaatt tttttttaac
24360 cagaattctg gcagtggccc ctgacttgcc tctgtgctcc cagccccgct
gtcctctgat 24420 ccatcctcca tgccagcctt tttcaatctg ctggtcacga
ttcattgatg ggttaggaaa 24480 tcaatggcat cacaactagc atttagaaaa
aggaaatagg cgttcccgcc gtggcacagc 24540 agaaataaat ccgactagga
accataaggt tgcgggttca acccctggcc ttgttcagtg 24600 ggttaaggat
ccggcattgc cgtgggctgt tttgtaagtc acagacatgg ctctgatccg 24660
gcattgctgt ggctctggcg taggcctgca gcatcagctc caattagacc cctatcctgg
24720 gagcctccat atgctgcaag tgcagcccta aaaaaaataa aaaaataaaa
aaaaataaat 24780 aaaagaagta gacaaattgt atagaacaac cctgagtatg
ttgcctgagc acatataaca 24840 agggtaagta ttatttcagg aaactctggt
ttcacagata ctcttggcat atggacccct 24900 agagtcctga tgtaaaatat
attcttcctg ggatcttagg caagaagttt gaaagctcca 24960 actctgcact
gctgccaaag aaatgatttt taagtgcaaa actcttcccg ttcccttccc 25020
tgtataaaat tccataggat ctctccagtg cctctaggat aaaggcagtt ttcattctct
25080 agttcaaggt gagagaagat tttaattatt tcacgtttta gtggggaatt
caagagtctg 25140 gcacctgaca tttgctgaac tctctccatt atccctctct
agttccccag acgcatccta 25200 tggtagaaat tcgcaaacta gagtgagcgt
cagagtaacc caaggaaact gggtaaatgc 25260 agctccctgg gctctacccc
ctgagattct gattcagtag atctgaagca gagccctgga 25320 atatgcatat
gcatcattgt gtcacaccaa gcattctggg taatgagagt tgatgttagg 25380
ttctcagtag taagacaagt atagagattc cgggggactg agtgctcagc tctgccttgg
25440 ggaggaggga gagggctaaa gagaacagga gatggggaca gggaatgctc
aacctccaat 25500 cttaggcatt tgagctatgt cttaggggtc aggaggaggt
taccaatata gtgattaaga 25560 gattgaggtt ccagtcagag ggatatgctg
gagaaggggg gtgaaaataa tgtcataggt 25620 ttggtgagtg cagatacttt
gagtttttta atatttttat tgaaatatag ttgatttaca 25680 atgctcttag
tgagtacaat tactttgaat aagtgcatag atgtatgcca ttcttccaga 25740
aatgatttat tgagctcctt tgggcatcat gctaagtaca ggggaaacag ctgtgaagag
25800 gtccttccct tatgaagtca ttcatcccct tcagtaaatg aaggtaaagg
aaaaggatga 25860 gacagggacg ccgtgttgga ccagggtcag aaaggcctta
taagaccttg cctggagggc 25920 aaggaacttg cctgtgagta aggagagctt
gagaaagcga taaagcaaag aaggaacatt 25980 actgcattgt gttttagaaa
aaccatgtcc tggggaagaa ctcctagagt caggggggcc 26040 agttgggaga
ctgtgctttt ttccaggagg agataagtga ggctgctggc tgagatggag 26100
caaggattta gagaagcaga tatgagattc atttagaagt tagacatttt aggatctgac
26160 acataattta tcaccaaaac cagtgcatct ctggctttgg gccaccagtt
ttggagaagt 26220 ggaatgtagg gacctaccat tacctgccaa tctttactac
acagatgcct atttccctcc 26280 tcatatttcc tttctccaga tcacgtccta
ttctattgcc aggactcaag attccacctt 26340 gcatgcagtg atccatcttc
acactggatg gacagctcta gggatgtcag agcacactct 26400 tgtccatact
gctgactggg tctcctgtca gcccatctgt ctatcagctg tggtattatt 26460
agtataataa gagggctgta tatgagagac acaaaattct aggtgtagct caaagatagg
26520 ctagagttat tcctatgtac aacaaatatt tatgggaccc cttctgtgta
ctgtcatggt 26580 tgctgctttc atcatacttg tagtctaatg gaggtggggg
cagggcagga ataagcggat 26640 gtccacaaaa tcagtaagac cacttatatt
caacattttc ataatttagt tatttgagcc 26700 caaagggtcc acatccgtgg
tattccaact tttttttccc cggacatgga tctttatctt 26760 tttttttttt
tcttttttgc ggccagacct gcggcatatg gaagttccca ggccaggggt 26820
tgaatgggag ttgcagctgc ctggtctaca ccacagccac agcaaggtgg gatctgagct
26880 gcatctgtga catacaccgc agctgaggta acaccagatt ctgaacccac
tgaatgaggc 26940 cagggatgga acccgtctcc ttatgaacac tatgtcatgt
tcttcaccct ctgagccaca 27000 acgggaactc cagacttcgt ctttaaatgt
attctgactt ggagagctat cacactaagc 27060 aattaacagg agctgacctg
gtttaggctg gggtggggcc ctactcctca atgttccctg 27120 aggcacatct
gtgggacccc tgggcatcat ctatctgagc agccttagag ctgctcatcc 27180
agttgactgt tgatgtagaa gtgcaaactt ctgccttcct tatttgttgc tttctttttt
27240 cattgttctc tcccctttgt gtctttaagc aagggcatcg tagagcctcc
agaagggact 27300 aagatttaca aggattcctg ggattttggc ccatatttga
atatcttgaa tgctgctata 27360 ggagatgaaa tatttcgtca ctcatcctgg
ataaaagaat acttcacttg ggctggattt 27420 aaggattata acctggtggt
cagggtatgc tatgaagtta ttatttgttt ttgttttctt 27480 gtattacaga
gctatatgaa aacctcttag tattccagtt ggtttctcaa taagcattca 27540
ttgagcctta ctgactgtca gacggagggc gtattggact atgtgctgaa acaatccttt
27600 gttgaaaatg tagggaatgt tgaaaatgta gggaatgaaa tgtagatcca
gctctgtttc 27660 tcttttggag gattcttttt cctccatcac cgtgtcttgg
ttcttgtttg ttttgggttt 27720 ttgtgggtgt tgtattgtgt tgtgttggtt
atggcagtga cagctattta aactgtgaaa 27780 cgggggagtt cccgtcgtgg
cgcagtggtt aacgaatccg actgggaacc atgaggttgc 27840 gggttcggtc
cctgcccttg ctcagtgggt taacgatccg gcgttgccgt gagctgtggt 27900
gtaggttgca gacacggctc ggatcccgcg ttgctgtggc tctagcgtag gccagcggct
27960 acagctccga ttggacccct agcctgggaa cctccatatg ccgcaggagc
ggcccaaaga 28020 aatagcaaaa agacaaaata aataaataaa taaataagta
agtaaaataa actgtgaaac 28080 ggggagttcc cttcatggct cagcagttaa
caaacccagc taggatccat gaggatgtag 28140 gttcgatccc tggccttgct
cagtgggtta agaatccagc gttgctgtga gctgtgatgt 28200 aggtcgcaga
tgcagcccag atcctgcatt gctgtggctg tggcgtaggc tggcagctga 28260
agctccgatt caacccctag cctgggaaca tccatatgct gcaggtgtgg ccttaagagg
28320 caaaaaaata aaaaaataaa aaataaataa attgtgggac agacaggtgg
ctccactgca 28380 gagctggtgt cctgtagcag cctggaagca ggtaaggtaa
ggactgcagc tgggtaagga 28440 ctgaattgca ccaactggga agtaagccta
gatctagaac ttaagttagc cctgacatag 28500 acacacagag ctcaccagct
aagtggttca gcttataagc tggtcactga aactgaggat 28560 gtccacaaaa
gcaaaataag tagcaacagg cagcgggatg caagagaaag aggaggccta 28620
aaatggtctg ggaatccctg ccatacctat attttatcct acttatattt agtgcctgaa
28680 tgtgtgcctg gagagcaaag tttagggaaa gcatcgggaa atgcacagta
ttcataccct 28740 taggaacaaa gatcagttac ctccagggta aagactattt
ccaagtttaa atttcaaccc 28800 ctgaacatta gtactgggta ccaggcaaca
cttgccatcc tcaaaatcaa tgaatcctaa 28860 aattcaacct gggggtcagt
gacagtctgt gacaaagttt ttgctggtca gtaacgaaat 28920 aagtatgagc
accatctgag tatggtcacc aagatgtcaa ctctctttcc tttggacgaa 28980
ttgtcattat tccaagatta ggtcctttct atttttgagg tgtgaaaaca tctttccttt
29040 cataaaataa aaggatagta ggtggaagaa ttttttttgt tttttggtct
ttttgctatt 29100 tctttgggcc gcttctgcag catatggagg ttcccaggcc
aggggtcgaa tcggagcttt 29160 agccaccggc ccacgccaga gccacagcaa
cacgggatcc aagccgcatc tgcagcctac 29220 accacagctc acggcaatgc
cggatcgtta acccactgag caagggcagg gaccgaaccc 29280 gcaacctcat
ggttcctagt cggattcgtt aaccactgcg ccacgacggg aactcctaat 29340
gatactcttt tatatttagc tactatgtga tgatgagaaa cagtccacat
tttattattt 29400 tttagccaat ttgatatctc attactaaga taatgataat
tttctctata aattttattt 29460 aagttagtgt tatgaagtgg ttttgctagt
gtagaaggct aggatttgaa ttcagttcaa 29520 gaaagaagag agggagggag
ggagagggat gggtagaggg atggggcagt gggagagagc 29580 aaagaggaga
gacagttttt gtattaattc tgcttcattg ctatcattta agggcacttg 29640
ggtcttgcac attctagaat tttctaagga ccttgaccgc cagattgata tgcttcttcc
29700 ctttaccatg ttgtcatttg aacagatgat tgagacagat gaggacttca
gccctttgcc 29760 tggaggatat gactatttgg ttgactttct ggatttatcc
tttccaaaag aaagaccaag 29820 ccgggaacat ccatatgagg aagtaagcag
gaataccagt ggaagtgccc ctttcttcct 29880 tccttcctaa ataaactttt
ttattttgga acaactttag agttacagaa aagttgcaaa 29940 gatattatag
acagtagtgt ttatatatat atataaattt ttttttgctt tttatgacca 30000
cacctgtggc atatggaggt tcccagtcta ggggttgaat tggagctaca gctgccagtc
30060 tgtgccataa ccacagcaat gcaggatctg ggccacgtct gtgacctaca
ccaaagctca 30120 cagctggatt cttaacccac tgagcaaggc cagggattga
acctgcatcc tcgtggttcc 30180 tagttggatt cgtttccgct ttgccgcaat
gggaactcca aattattgtt aatatcttac 30240 tttactgggg tacatttgtt
acaaccaata ctctgatact gaaacattac tgttaactcc 30300 gtacttgctt
ctttttgagt catttgcaaa gactggcttc ttgacctgct tccttccaaa 30360
cagctggcct gcctatgctg ttctcagacc tgcaagcact gatctctgcc ccccttgcct
30420 tctctccagt ggtgtctcct tccccaaaca aacccagtgt ggctctggaa
agggagttaa 30480 gtcaacataa accaacacat attttgttga gctccaattt
tgagcaaatc cctcacctac 30540 ggcagacagg catgatgtta agaactaggg
ctttggacac aaggtcaaga ccaagaaggg 30600 ttcctcaccc ctactgattc
agataaccaa taatgaggct ttgaatccct gtccaaaggt 30660 tgtttttttt
cccttctatt gagcttcttg ccaccttatc agtttttttt atgacagtca 30720
aatgacatga tatatgtgag catacatggt aatttttaat tctatataaa tgaatcacta
30780 aataaattag gaggatatat agtccacctt taagcgtatt acacgtgtca
catgaatgtg 30840 tggcgactta attgtagagg tttaaatgta gcttcctata
atagatgtgt tcctaaacta 30900 cattttaatc attggacttg tatttttatg
ttagcacttg ctgttgaaga aaagcctatg 30960 ccaaaagttc agtgaaacca
ataatccact gccagctttc tgagttaaaa aaaatccctg 31020 ggttttcaca
cacaggaaca ccctgtgtga aacactcatt tagagcaaaa tgcatctgat 31080
aaggagttcc tgttgtgcct caactggtta aggacctgac attctccatg agaatgtgag
31140 tttgatcccc ggccccactc gatgggttaa ggatctggtg ttgccacaaa
ctgcagctcc 31200 gattcatctc ctagcctaga aacttccaca gcccagaata
tgccacagaa ttcggctgtt 31260 taaaaaaaaa aagaaaaaaa aaagaatcat
aaatgtgttg gtttgttcac caaatacatg 31320 ataacttgct cttgccaagc
tcagcttcat aaatattaag tcatttaata cagcagccac 31380 cttatgaaca
gatattacta tacttcccat ttacagataa ggaaaatgcc atatttaacc 31440
aagagattaa ataactttcc cgaggtctta tagcaagtaa atcatggtgc aggggtttga
31500 ccacacgcag tctatctcca gagtctgtgt atttagccac tgttttactt
tcaaatttaa 31560 atttataaaa cttctaaatt atctgttaac cataatcttt
ggaattttta aaaccacgag 31620 ttcctataaa atgtttcatt gaaagtaagt
cacttttcca tagcttttga taatacatct 31680 gtaggataaa gtaagccaca
gctctcttgc agacttggta caccctgggg caaagcatca 31740 tgcctgtcac
gtacatggtg gtccttactt tgactctcag tgcttttatt gcccaggaat 31800
tttgtgagat ttctagttgt tgaggtttgt ttaaagaggt tatgccggta cttggaagag
31860 ctcttttctt gctacctgga gccttctcat atttcctttt tgaggaggga
catgaattgc 31920 ctttcaaact cataaatata ttttctagta cacaagtctc
catcttcctt agacgcatgg 31980 ctcctggagt tctccatcct cctgctccac
tttgggtggg ctcctctctg ggtctgccac 32040 caatctgcca cccagagaca
tccttgaccc acttccagac cccaccatgg cttcactttc 32100 ttcgctttcc
tcctttgtgg aaccttctgc ttaagaatct gaggaagaaa atttgcacgt 32160
gagctaaact ggaggtactt tcctgcctgg tcttgcacga tagcttggct gagcccatga
32220 tgctgggtgg ctgttacttt ccatggacac ccgaaggcgt tgctcctttg
gcttctagtt 32280 gcatgcagtg ttgcttatcc caggctgatc tttcttccac
tgtaggtgac ttttaagaat 32340 taagggatta atctatatct acaacaacaa
caacaaagac cttttcaagc tgaggtaggg 32400 ctttctgtat atgtttggag
tggttatcca gcagacttta cttgaaggca ggggtcatat 32460 cctcaagtgc
tcataaacgg accacagaaa gatctcataa ttgggtggag ctgggtgggg 32520
accgtgtcat gtggccagga aatgccagat gggaagggag tggcccttac tgagctccag
32580 ctgaactctg aattttctag aaaactcaga aatctggatt tttcatgtgt
aatacccaga 32640 tttatagatg tggaaagcta attctttttt tttttaaggg
actataggca atgaactaag 32700 atctaggttg tatttggaca aggggtcatc
agtttaagct gtgtagttga gcgctcagct 32760 attgggctga gggaccccta
aatactgaga cggggaggtc cttgctctgg ggcatcacaa 32820 gtacactccc
tggtctcatt caaacacttt tcctacaaaa ttgatcccat ttcttcagtg 32880
cactgtctga atgcatttgg cccagagccg tgctgaggca tagggaaggg gtccacggtt
32940 tcatggcatc gttttgtgct gtgtgtccct gctgtcgtcc aggataccta
cctctcctcc 33000 tcctgcatct gaatgtcccc ccacagactc tctgggattc
tacagcctct ggcctgttcc 33060 tcagacacct cttacctgcc agctttccag
attcacatta gttagtccaa atctactgcc 33120 gtcagtgact cacttcattt
cttcttctcc gaggcagttc agcccggtac agttgttttg 33180 tcaacacttc
agttgagtct ggaagatgtg catgggttat gcacgagagc ggtccatcat 33240
tttgagctag aagtcctttc tcagcccaga gacaagtcct catctccttt acttcctgac
33300 tcttcttcct ctgcatcctt ccaagatatc tctttctcca gccaccacct
aaatctcttc 33360 ttttcccggg gttccgtgct caacccactc ttcttcttaa
atctgtggct gggtgaacgc 33420 atctgctggc accacttctc tgctaaagac
tccaaaaatc cataggtcct gcccggcctt 33480 tgcccacctc tctccaacac
tgtccagctt tagatgtaga gctaatcccc ccagagatat 33540 cattccctgg
atgtctaagt cctttggtat ctcactttca gcgtgttcaa aatcctctta 33600
caactgttct ttctcctttt ccatcttgat tattggcaac atgccagcct ttcccctacc
33660 cccagcagtg agccaagcta gaaacaaggg cttaatcttc aatctttcct
tctccatccc 33720 taaacctaat gagtctccaa gcccttccca gtttacaccc
taaatgttgc tcaaaacatc 33780 ccctagttct tccacgtgct ctcctctata
ttgaaaggtc aagaaaggcc atcttccctc 33840 cactgtgagg aaatagatct
tgatactgcc cctgagctgg gcagtcctcg acctgacaaa 33900 ctgtgcagtg
tttctaaatc tctactggca aaatgagagt gcctttgacc tgtgttgcga 33960
tctcagatca cagtggatgt aattgtttta taggaatggt gaacgaaaaa gaagtaaatc
34020 cctaatgcca aactcctgat cattctatgt catttaatag cctgtcattt
atgataaagt 34080 ttcctctact ggcattagca caatacttct caggaaaaaa
aaatatgatg ccagatactg 34140 aaaagctcct gggtaaacat gaacatgggt
accgataaaa tggtgaagcc agtccaatct 34200 tagagtgact tcccttcatg
ctacttcatg ctcttttttt tttttttttt taagaaaaac 34260 cccttttttt
tttctcacac cagtcacaga ggagaccgag gcttagcaag gttaaggtca 34320
catgattagt aagtgctggg ctgaaactca aaaccatctc tgcttgtctc ctaaccctgt
34380 gcacctctga ctattcaaca gatcctgtgt caggagttgg gattctttga
aggtaagggc 34440 cttgaccacc gaattaaggt aatcttgctc tgtggcaggc
cttgttttca gtattttaag 34500 tacactggct caggtaatcc tcacaacagc
cccaggagga atgttctatt acctccactg 34560 tatagatgag gaacttgagg
cacagaatgg ttgccaaggt cacacagcta tattgggggt 34620 tcatacccag
ccatccaact ctgtctgtac tctctgccac tctgcacccc cagctcctga 34680
tccacttcct gtttccatcc ctcgatttct gctgcactca ggggcccctc tccccctcgg
34740 cctgtgagat ctgcttcagt aggcttttct ccctgactcc tccatccctg
tccttacagg 34800 cagctgcttc tctccgggac acgaggggtc catacggaca
ctctctactg gctgggttgc 34860 gcctaactcg tgattcctcc tctgtttcag
attcggagcc gggttgatgt catcagacac 34920 gtggtaaaga atggtctgct
ctgggatgac ttgtacatag gattccaaac ccggcttcag 34980 cgggatcctg
atatatacca tcatctgtaa gtccgaaaat gcctgtcgtg tgtgccttag 35040
gctgctgcgg aggaggccag ggctatataa gcagagtcag tgactgactg tgccctgcag
35100 tgttgatggc catggagatt ccaccgttag agcttttttc tttgttaacc
ttgaaggcaa 35160 atctggttag gaagataact ttcaaagagt caccatctgg
acattcatgc ccatgtgctt 35220 caatcctgta tacaagcagt ttagagtaca
gggaagggaa ggacattatg aaagggagag 35280 ggtgtgtttg gatccagcag
ctccatcctc agaatttatc tgaagacact gcaaaattac 35340 taagaatcac
tatgacaaga atgaggatgg ggtgatatgg caaagttgtg atcctggaag 35400
accttcatct cccatgttgc ccaactctga acatgaattt ggtgaactag ttggttaagg
35460 ggatgatcct ccaagtttct ccctggttga gctccaaaaa ccatgtaagt
ttctcatagc 35520 aaaaccgtat aggtccttag ggctttagtt ggaatatttg
tgctgaaatg ctggaaagcc 35580 ccatttgcca tttttgtatt tgcaaaataa
tcatcaagag gggagaatgc attctttcat 35640 gaccactgac cctctgaaaa
ggtcaggaat ttagtctgaa gtaggcaagc ctcctacccc 35700 gcttctgcca
tgagcttgca cgcacaggcc tgtcttgaca tttcttcttt atagatttct 35760
ttttgaatat cttgaaattg ctttaaaaat atttaaagaa tgtagaatta tataaaataa
35820 aaaggaaata accccacacc tcccacaaaa ccctgtttcc tgcctttctc
cacccactct 35880 ccagggtaac acttggtaac agcatagttg tatcacccca
ggcctatttt tgagcatatc 35940 agcatttcaa gaaatgtatt ttttctcaat
aaaacatccc ttatagttga ggaggggagg 36000 ttatcattcc tgggttttgt
tttttttttt tttttaatgt aatcctggta catcggtaat 36060 ttgcattttt
tattcattaa tatctttggt atttctagtg ttgggacaca caggtcaacc 36120
tcagtttttg ggtttttttt tttgtctttt tgtctttcta gggccacacc tgcagcatat
36180 ggacgttccc aagctaggag tctaatcaga gctgtagcca ccagcctacg
tcatagccat 36240 agcaacgtca gatccaagcc gtgtctgtga cctacaagca
cagctcatgg caacaccgga 36300 tccttaacca ctgaacgagg ccaggggatc
gaacacacat cctcatggat cctagtcatg 36360 ttcattaacc actgagtcat
gatgggaact ccaacttcaa ctattttaat gtctgtaaaa 36420 cattccattt
ggaaaccatt tcatttgtaa agcaaaatga aaacattttg ttcattttca 36480
acagagttcg tagctgactt ctgttctgga aaaaaggaaa tggagcaaat ttgagtgaga
36540 aagattcaaa gataactttt cttttaaaaa aaattatatc ttggaaactt
ctgggctatt 36600 gattctgaag actatttttc tatatactgt tttgatagca
aagttcataa atgtgaaagg 36660 atcctgcgat gaatcttggg aagcagtcat
agcccaatat atctttgttg cttttaaaat 36720 gagatttagt ttactaaata
tttttctgat cataaaaata acacagatct accgcagaaa 36780 atttggaaaa
aaaaaaactt ttaaattcaa aaaacagtta aaccacaaat gatcccacca 36840
tccagagagc aatttgtact ttggtgtcta gttcatcttt ctttttctgt ttacaagcac
36900 atataccaca agcatttttt caaaaaatga aaatgggata atactataca
tacgtctgta 36960 cacctgcata gttactgaac agtctttgat ctaccctgta
agtttctaac ttttcattat 37020 ttgaaatgat gttttggcaa agaaatatgt
aggtgtgtct cgcacacttt cataatgatt 37080 tcttaggata aatttcttag
gataaattca taatgatttc ttataataat ccatactctg 37140 ccaactgatc
ttcagggaag ccaactcgcc ttctcagaaa taacatataa cccatttact 37200
tgccctctca ccaatactag gtcctaatgt ttttgtgtac agattctata tttttacata
37260 caagaattcc ttaaagcaag gcatgtcaca gaaaaataga aggaagacac
aattgtcatg 37320 tttaaggact gcattctgta ccaaaaatgc taagttaaat
gaacatctga aacagtacag 37380 aaacgctatc tttcagggaa agctgagtac
caggtactga acagattttg gcaaatacag 37440 caggcatgga tgtttccaaa
acatgttttt ctactttatc tcttacaggt tttggaatca 37500 ttttcaaata
aaactccccc tcacaccacc tgactggaag tccttcctga tgtgctctgg 37560
gtagagagga cctgagctgt cccaggtaaa gcatcctgca ggtctgggag acactcttat
37620 tctccagccc atcacactgt gtttggcatc agaattaagc aggcactatg
cctatcagaa 37680 aacctgactt ttgggggaat gaaagaagct aacattacaa
gaatgtctgt gtttaaaaat 37740 aagtcaataa gggagttccc atcgtggctc
agtggtaacg aaccctacta gtatccattg 37800 aggacacagg ttcaatatct
ggcctcactc agtcggctaa ggatccagtg atgccgtgag 37860 ctgcagtgta
ggccacagac gtggctcaga tctggtgctg ctgtggctat ggtgtaggcc 37920
ggccccctgt aactccaatt cgacccctag gctgggaacc taaaaagacc ccaaaaaagt
37980 cgctttaatg aatagtgaat acatccagcc caaagtccac agactctttg
gtctggttgt 38040 ggcaaacata cagccagtta acaaacaaga caaaaattat
cctaggtggt cagtgggggt 38100 tcagagctga atcctgaaca ctggaaggaa
aacagcaacc aaatccaaat actgtatggt 38160 tttgcttata tgtagaatct
aaattcaaag caaatgagca aaccaattga aacagttatg 38220 gaagacaagc
aggtggttgt caggggggag ataaggggag gcaggaaaga cctgggcgag 38280
ggagattaag aggtaccaac tttcagttgc aaaacaaatg agtcaccagt atgaaatgtg
38340 caatgtggga aatacaggcc ataactttat aatctctttt ttttttttgt
cttttttgcc 38400 ttttctaagg ctgctcccgt ggcatatgga ggttcccagg
ctaggagtcc aaacagagct 38460 gtagctgcca gcctacacca gagccacagc
aacacgggaa ccttaacccg ctgagcaagg 38520 ccagggatcg aacccgagtc
ctcacagatg ccagtagggt tcattaacca ctgagccacg 38580 acaggaattc
cagggtctgt tgtgttctta aaacacttcc aggagagtga gtggtatgtc 38640
ataagtaaac aataaatgtt aaccacaaca agcttatgaa ataaacagga aagccatatg
38700 acctacaatc agtcattggg agaatccaca aaaggttgag cagaggatca
attccagctc 38760 acactccagt tttagattct cccctgcctt aaagcatcac
agactacata atctgagctg 38820 aagaataaaa attaaaactc accccagtgc
aaaacagaaa tgaaaaagta ttaaaacgag 38880 gttcatactg ttgttcatta
gcaatatctt ttattcacag gggtgcccaa caacatgaaa 38940 aaatcaagaa
tttattgctg ctacgtcaaa gcttatacca gagattatgc cttatagaca 39000
ttagcaatgg ataattatat gttgcacttg tgaaatgtgc acatatcctg tttatgaatc
39060 accacatagc cagattatca atattttact tatttcgtaa aaaatccaca
attttccata 39120 acagaatcaa cgtgtgcaat aggaacaaga ttgctatgga
aaacgagggt aacaggagga 39180 gatattaatc caagcataga agaaatagac
aaatgagggg ccataagggg aatataggg 39239 49 14929 DNA Porcine 49
tctaatgcct tgtggaagca aatgagccac agaagctgaa ggaaaaacca ccattctttc
60 ttaatacctg gagagaggca acgacagact atgagcaggc aagtgagagg
gggctttagc 120 tgtcagggaa ggcggagata aacccttgat gggtaggatg
gccattgaaa ggaggggaga 180 aatttgcccc agcaggtagc caccaagctt
ggggacttgg agggagggct ttcaaacgta 240 ttttcataaa aaagacctgt
ggagctgtca atgctcaggg attctctctt aaaatctaac 300 agtattaatc
tgctaaaaca tttgcctttt catagcatcg aacaaacgac ggagatcctg 360
ttgtgcctct cacctgccga agctgccaat ctcaaggaag gaatcaattt tgttcgaaat
420 aagagcactg gcaaggatta catcttattt aagaataaga gccgcctgaa
ggcatgtaag 480 aacatgtgca agcaccaagg aggcctcttc attaaagaca
ttgaggatct aaatggaagg 540 tactgagaat cctttgcttt ctccctggcg
atcctttctc ccaattaggt ttggcaggaa 600 atgtgctcat tgagaaattt
taaatgatcc aatcaacatg ctatttcccc cagcacatgc 660 ctaacttttt
cttaagctcc tttacggcag ctctctgatt ttgatttatg accttgactt 720
aatttcccat cctctctgaa gaactattgt ttaaaatgta ttcctagttg ataaacagtg
780 aaacttctaa ggcacatgtg tgtgtgtgtg tgtgtgtgtg tgtgtttacc
agcttttata 840 ttcaaagact caagcctctt ttggatttcc tttcctgctc
tctcagaagt gtgtgtgtga 900 ggtgagtgct tgtccaaaca ctgccctaga
acagagagac tttccctgat gaaaacccga 960 aaaatggcag agctctagct
gcacctggcc tcaacagcgg ctcttctgat catttcttgg 1020 aagaacgagt
gctggtaccc cttttcccca gccccttgat taaacctgca tatcgcttgc 1080
ctccccatct caggagcaat tctaggaggg agggtgggct ttcttttcag gattgacaaa
1140 gctacccagc ttgcaaacca gggggatctg gggggggggt ttgcacctga
tgctccccca 1200 ctgataatga atgagggatt gaccccatct tttcaagctt
tgcttcagcc taacttgact 1260 ctcgtagtgt ttcagccgtt tccatattag
gcttgtcttc caccgtgtcg tgtcgtcaat 1320 cttatttctc aggtcatctg
tgggcagttt agtgcgaatg gactcagagg taactggtag 1380 ctgtccaaga
gctccctgct ctaactgtat agaagatcac cacccaagtc tggaatcttc 1440
ttacactggc ccacagactt gcatcactgc atacttagct tcagggccca gctcccaggt
1500 taagtgctgt catacctgta gcttgcttgg ctctgcagat agggttgcta
gattaggcaa 1560 atagagggtg cccagtcaaa tttgcatttc agataaacaa
cgaatatatt tttagttaga 1620 tatgtttcag gcactgcatg ggacatactt
ttggtaggca gcctactctg gaagaacctc 1680 ttggttgttt gctgacagac
tgcttttgag tcccttgcat cttctgggtg gtttcaagtt 1740 agggagacct
cagccatagg ttgttctgtc accaagaagc ttctgcaagc acgtgcaggc 1800
cttgaggtct tccgacttgt ggcccgggga ctctgctttt tctctgtcct tttttctcct
1860 tagtgggcca tgtcctgtgg tgttgtctta gccagttgtt taagggagtg
ttgcagcttt 1920 atgattaaga gcatggtctt tccttgcaaa ctgcttggtt
tagaagcctg gctccaccac 1980 ttagcggctc tgtgacctcg gacacatttc
ttagcctttc tgggcctcgc tcttcttcct 2040 cataaagtga aaatgaaagt
agacaaagcc ttctctgtct ggctactgag aggatggagt 2100 gatttcatac
acataaagca cttaaaataa tgtctggcat atgatacatg ctcaataaat 2160
gtcacttaca tttgctatta ttattactct gccatgatct tgtgtagctt aagaacagag
2220 gtctttacag gaattcaggc tgttcttgaa tctggcttgc tcagcttaat
atggtaattg 2280 ctttgccaca gactggtctt cctctccttc acccaaagcc
ttagggggtg aacgatccca 2340 gtttcaacct attctgttgg caggctaaca
tggagatggc accatcttag ctctgctgca 2400 ggtggggagc cagattcacc
cagctttgct cccagataca gctccccaag catttatatg 2460 ctgaaactcc
atcccaagag cagtctacat ggtacactcc cccatccatc tctccaaatt 2520
tggctgcttc tacttaggct ctctgtgcag caattcacct gaaatatctc ttccacgata
2580 cagtcaaggg cagtgaccta cctgttccac cttcccttcc tcagccattt
ttcttctttg 2640 tacataatca agatcaggaa ctctcataag ctgtggtcct
cattttgtca atctaatttc 2700 acagcctctt ggcacatgaa gctgtcctct
ctctcctttc tgcctactgc ccatgagcag 2760 ttgtgacact gccacatttc
tcctttaacg acccagcctg ctgaatagct gcatttggaa 2820 tgttttcaat
ttttgttaat ttatttattt catctttttt tttttttttt tttttttttt 2880
tttttagggc cgcacccatg ggatatggag gttcccaggc tagggatcca atgggagctg
2940 tagctgctgg cctacaccac agccacagca atgcacaatt cgagccaatc
tttgacctac 3000 accagagctc acggcaacac tggattctta acccactgat
tgaggccagg gatcaaactc 3060 tcgtcctcat agatacgagt cagattcgtt
aacctctgag ccatgatagt tgttagttac 3120 tcattgatga gaaaggaagt
gtcacaaaat atcctccata agtcgaagtt tgaatatgtt 3180 ttctgccttg
ttactagaaa agagcattaa aaattcttga ttggaatgaa gcttggaaaa 3240
aatcagcata gtttactgat atataagtga aaatagacct tgttagttta aaccatctga
3300 tatttctggt ggaagacata tttgtctgta aaaaaaaaaa atcttgaacc
tgtttaaaaa 3360 aaaaacttga ctggaaacac taccaaaata tgggagttcc
tactgggaca cagcagaaat 3420 gaatctaact agtatccatg aggacacagg
tttgatgcct ggcctcgcta agtgggttaa 3480 ggatatggtg ttgctgcagc
tccaattcaa cccctatcct gggaaccccc atatgccacc 3540 ctaaaaagca
aaaagaaagg tgctgcccta aaaagcaaaa agaaagaaag aaagacagcc 3600
agacagacta ccaaatatgg agaggaaatg gaacttttag gccctatctc caactatcac
3660 atccctatca ccgtctggta agaaatggaa aaaatattac taagcctcct
ttgttgctac 3720 aattaatctg attctcattc tgaagcagtg ttgccagagt
taacaaataa aaatgcaaag 3780 ctgggtagtt aaatttgaat tacagataaa
caaattttca gtatatgttc aatatcgtgt 3840 aagacgtttt aaaataattt
tttatttatc tgaaatttat atttttcctg tattttatct 3900 ggcaaccatg
atcagaaatc tttaaacaat caggaagtct tttttcttag acaaatgaaa 3960
atttgagttg atcttaggtt tagtacacta tactaggggc caagggttat agtgtgacta
4020 ttaaatcaca gataatcttt attactacat tatttcctta tactggcccc
acttggatct 4080 tacccagctt agcttttgta tgagagtcat ccttaaagat
gactttattc tttaaaaaaa 4140 aaaacaaatt ttaagggctg cacccatagc
atatagaagt tcctaggcta gcggtcaaat 4200 tagagctgca gctgccagcc
tatgccacag ccacagcaat gccagatctg agctgcatct 4260 gtgacctaca
ctgcagcttg cagcaatgct ggatccttaa cccattgaac aatgccaggg 4320
attgaacaca catcctcatg gatactgctc aggttcctaa cctgctgagc cacagttgga
4380 actccaaagc agactttatt ctgatggctc tgctgatctc taacacgtta
ttttgtgcca 4440 tggtgtttat cttcacttta ctcaagtcag ggaaacacga
agagtctcat acaggataaa 4500 cccaaggaga aatgtgcaaa gtcacataca
aatcaaactg acaaaaatca aatacaagga 4560 aaaaatatct tcactttcaa
aatcacctac tgatgatgag tttatatttc cttggatatt 4620 tgaatattag
ctattttttt cctttcatga gttttgtgtt caaccaacta cagtcgttta 4680
ctttgatcac agaataatgc atttaagcct taaatagatt aatatttatt ttcaccattt
4740 cataaaccta agtacaattt ccatccaggt ctgttaaatg cacaaaacac
aactggaagt 4800 tagatgtaag cagcatgaag tatatcaatc ctcctggaag
cttctgtcaa gacgaactgg 4860 gtaaatacca tcaatactga tcaatgtttt
ctgctgttac tgtcattggg gtccctcttg 4920 tcaacttgtt tccaatctca
ttagaagcct tggatgcatt ctgattttaa actgaggtat 4980 tttaaaagta
accatcactg aaaattctag gcaagttttc tctaaaaaat cccttcattc 5040
attcatttgt tcagtaagta tttgatgaga ccttaccatg tgtaaacatt gcactaggta
5100 ttaagaaata caaagatgga taagatagag tcggcgtaaa tgagatgata
taatgagacg 5160 ttataatgaa actcacaatt
ccagttggga aataaagtcc ttcaaattcc atgactcttt 5220 ctggcacacg
ttagaggcta cagcttctgt gtgattctca tgctggctcc acttccactt 5280
tttccttctt cctactcaag aaagcctata gaaatatgag taagaagggc ttaatcatag
5340 gaataaattt gtctctgttc taagtgatta aaaatgtctt tatcagtata
aaaagttact 5400 tgggaagatt cttaaaactg cttttacaca ctgttctaga
atgactgtta tataaataaa 5460 aaagtagatt tgatctaaca caattaaatg
acctttggaa atattgacta attctcacct 5520 tgcccctcaa agggatgcct
gaaccatttc cttcttttgc cagaaagccc ccaccctttg 5580 tctgttgacc
tagcctagga aatcttcaga tcacgttgtt agcacgaact ggttacatgt 5640
gctgtacaaa tactatttaa ttcatctgat taaaaaaaaa gagataagaa gcaaaagttt
5700 gactatctta aactgtttgc gtaggtgaga ggacaattga ccatctactt
tatgagtatg 5760 taacccagaa acttaaagct ccttaaggga gctaagtctt
ttggataaga cctatagtga 5820 gaccttttag caaaatggtt aagactgaat
ggagctcact agcgtgggtt catatcctga 5880 tgctcaaaca cgcaattaaa
tgactttagg tgggttagtc tctgttcctt agtttcctca 5940 atgggagata
atattggtag tagcgatttt actgggttgt tgaaagaaca tctgttaaat 6000
gttcagaacg tgttacgaca gagtacagag taatgatttg cttgtatatg tatgactcaa
6060 atagtctgcc atatgccttg tgactgggtc ctgtggagca ggaaggaggg
atttcccacc 6120 cagcagaaag ttgggtaaac tggaaaatag actgaggcca
ggaaatgatg caaagcgttg 6180 atgttcactg ccacggcagg tgaagggcag
ggccagagtt gtcagtaggg tcaggggagg 6240 actggaaata accaagaccc
actgcacttt tcagcctttg ctccagtaag gtaatgttgt 6300 gagagtagaa
aattttgtta acagaaccca cttttcagta cagtgctacc aatactgtag 6360
tgatttcata ccacatccca agaaagaaaa agatggctca atcccatgtg agctgagatt
6420 atttggtttt attgttaaat aaatagcatt gtgtggtcat cattaaaaaa
ggtagatgtt 6480 aggaaagtag aaggaagaag actctcacct acattttcat
cactgttttg gtatctgcca 6540 gttgtcacct tggtcccctt ccccgcctct
cccctgcctc ctcttcctcc ttctcctttt 6600 tttggaatac aattcaggta
ccataaaatt taccctttta gagtgtttga ctcaatggtt 6660 tttagtattt
tcacatgttg tgctattact atcactatat aattccaggt cattcacatc 6720
accccccaaa gaaaccttct aactattagc agtccattcc cttcttccct cagcccctgg
6780 caaccactaa tctacttact gtctccatgg atgttcctat attgaatcaa
gctagcataa 6840 accccacttg ctcatggtca taattctttt ttatagtgct
aaattacatt tgctaatatt 6900 caattaagga tttctatgtc catattcata
aggaatattg gtgtgtagtt ttctctttgt 6960 gtgatatctt tgtctggttg
ggggatcaga gtaataatta ctgctctcat agaatgaatt 7020 gagaagtgtt
ccctcctttt ctatttattg gaagagtttg tgaagtatat tggtattgat 7080
tcttctttaa acatttggtc agattcacca gtgaagccat ctgggccatg gctaatcttt
7140 gtgaaaagtt ttttgattac taattaaatc tctttaattt gttatgggtc
tgctcctcag 7200 acgttctagt tcttcttgag tcagttttgt tcatttgttt
cttcctagga ctttctccct 7260 ttcatttgga ttatttagat tgatagtaat
atcccccttt taattcctgg ctgtagtaat 7320 ttgggtcttt tctctttttt
cttggtcagt ttagctaaag gtttgtaatt gtattaatct 7380 tttcaaataa
ctaacttttt tgttttgttt gttttttgtt ttttgttttt tgttttttgt 7440
ttttttttgc tttttaaggc tgcacctgag gcatatggaa gttctcaggc tagaggtcta
7500 atcggagcta cagctgctgg cctataccac aaccatagca atgccagatt
caagctgcat 7560 ctgcgaccta caccacaact cggccaggga tcacacccgc
aacctcatgg ttcctagtcg 7620 gatttgttaa ccactgtgcc acgacgggaa
ctcccgccca ttttttttaa cacctcatac 7680 tttaacataa agatgggctt
cacatggact gatagctcaa atgaggaagg taagactatg 7740 aaagtaatgg
aagaaatgta gactattttt gtgacctaga gattactgat acttcttgac 7800
ttttcaaaca atacttcaaa agtacagccc aaagggaaaa aagaaagaaa aaagaaacac
7860 acatatacac aaacctagtg aataagatat catcgataca ctacagattt
ctatgaactg 7920 gaagacccca tggacaaagt taaagaacat atgatagttt
gagtgattat tttgcaatat 7980 ttacaaccaa tgagggaata ttatccagct
tataggagga agtaatgcaa atcgacaaga 8040 aaaagatagg aaacccaata
taaaaattaa gaaaatacaa aaattaagaa aggatatgaa 8100 ctagcatttt
acaaaagaaa aatctccaaa agtcaatcag cacatgaaaa tatgctcaaa 8160
cctattaatt attagaaaac tacagactga agcaatgagg tgctttactt tacatctttt
8220 tgactgataa aaagttagaa acaaaggtga tatcaaatgt cagggataaa
aggatataga 8280 aatcgtcatg cctgtggtgg gagtatggcc ggtgcagtca
tgtgggaagg taatctgaca 8340 gtggttaggc agagcaggtt tatgaataca
ctgtggccca tcaatcccac gcctgtttat 8400 gtaccaaaga aatcctgttg
tggcagaatc tatgggtcca cccctgggag catgaattaa 8460 taaaatgtgg
caccagggtg tgtgaaactc cagctagaga tgagatgtcc acatggcaac 8520
atgaatgcat cttagaaaca tagatttgag tgaaaaagag taagaaacag ccgggaaacc
8580 caataccatt tataaaaatt aaagatgcac acatacaatg tagtaaatat
tttgcatgaa 8640 ctttcaaatg gttgcctaca gggggggaga gtaaagaaga
gtagaaaaca aagataaagg 8700 gagtaagtaa gtagctctgc ctggactgaa
tataatgtgt catgaactga gaaatatggt 8760 taacataatc ctcttaactt
gaggtcctaa atgaatgaat gagtccacta ttcatttacc 8820 cattctttaa
tgtgtattgc attataatcc atttttttag aaccaacgaa ttttgttccc 8880
ataactacta atcagcctgc cttttctccc tcattccctt atcagctcag gggcattcct
8940 agtttttcaa acgttcctca tttgaaccaa aaatagcatc attgtttaaa
ttatacttgt 9000 tttcaaatac gatgcttata tattccaagt gtgtttgccc
attttcttag gtggtagaaa 9060 tttttcattc tacttttcta tctactcaga
ttttcccgtt ggaattattt ccattgctat 9120 taaacttaga agtcccccct
gtgatatgcc atttttttca tactttttaa gcacttggtt 9180 gcttttcttt
gtgtctttaa gcacctagaa tacttataac cattgcacag cactgtgtat 9240
caggcagccc ttcctcttcc actaatttat ggtccttctc ttagactata ttaaactgtt
9300 atttaattag gatcctctct tcgtccttat gatttaatta ttatagtttt
ctaatatgtt 9360 tttattataa ttcctcttca ttattcctcc ctattaaaaa
ttttaatgaa ttccatttgt 9420 ttgttcttct agttaaatat taagtcataa
tccaaataac ttagatgtca ttagtttatg 9480 tggtcaaagt aaggatacca
catctttata gatgcaggca gttggcagat gtcatgattt 9540 tcttcagtgc
ataaatgcaa tttatctttg agcaaggggc ataaaaactt ttatggtatt 9600
ggctttgaaa taatagttaa gaactgcaga ctcagttttt cctgcttttc ttgaaaaaga
9660 acacttctaa agaaggaaaa tccttaagca tggatatcga tgtaattttc
tgaaagtctc 9720 ctgtaattcc ttgggatttt tgttgttgtt tgttggtcgg
tttttttggg tttttgtttg 9780 tttgttttgt tttgttttgt tttgctttta
gggctgcacc tgtggcatat ggaagttccc 9840 aggctagggg tccaactgga
gctacagctg ccagcctact ccacagccac agcaacatgg 9900 gatcctagct
gcatctgtga cctaaccaca gctcttggta atgccagatt gttaacccac 9960
tgagcaatgc cagagatcga atctgcctcc tcatggacac tagtcagatt agtttctgct
10020 gagccacaat gggaattccc aattccttgt atttttgaac tggttatgtg
ctagcatata 10080 attttgtttc ttgaatcttt gtgggttttt tttttttttt
ttttttgtct cttgtctttt 10140 taaggctgca cccacagcat atggaggttc
ccaggctaga ggtcaaattg gagctacagc 10200 tgccagccta cacaacaact
gcagcaaagt ggggcccaac ttatatgaca gttcgtggca 10260 atgccggatt
cctaacccac tgagcagggc cagggatcga acctgagttt ccagtcagtt 10320
tcgttaacca ctgagccatg atagtaactc ctgtttgttc agtcttgaac ctccttttta
10380 attctttatt ccttgagggt gaaataattg ccataataat actatcattt
attacatgcc 10440 ttctctgtgc taggcatagt gacactttag gatttattat
atcacttaat ccctacaaca 10500 actctgcaaa gtatgtatca taatcctatt
tgacagatca ggaaattgca gcccaggatg 10560 cagataatat gcatccatca
caagtgacta gatatagtcc ctctgctatt cagcagggtc 10620 tcattgcctt
tccattccaa atgcaatagt ttgcatctat tgtatatgtg ttttggggtt 10680
tttttgtctt tttttttttt tttgtctttt ctggggcctc acccttggca taggtaggtt
10740 cccaggctag gggtcaaatt gaagctgcag ctgccagcct acaccacagc
cacagcaact 10800 cgggatctga gcctcatctg caacctacac caaagctcac
ggcaacaccg gatccttaac 10860 ccactgagtg aggccagaga tcaaaccggc
aacctcatgg ttcctagtcg gattcattaa 10920 ccactgagcc acgatgggaa
ctccctaaat gcaatagttt gctctattaa ccccaaactc 10980 ccagtccatc
ccactccctc ctcctccctc ttggcaacca caagtctgtt ctccatgtcc 11040
atgattttct tttctgggga aagtttcatt tgtgccattt ttcattttac gggtaatttt
11100 tacttcagtt tcttccacta gcagttgtct taaagtgagt ataattaata
ttcatttgga 11160 aaatgtaagc aaaacatttt ttaaagggcc atgcccacag
catatgaaag tttctgggcc 11220 aggggttgaa tccaggctcc aagttgcagc
tgtgccctac actgcagctg ggcaatgctg 11280 gatcctttaa cccactgtgc
ccggctaggg atcaaacctg catttccaca gctacccgag 11340 ccattgcagt
tggattctta acccactgca ctacagtggg aactcccaca aaacattttt 11400
taatgtcctt tgaataaagt aggaaagtgc tcgtctttga gggcagggcg gcaatgccat
11460 ttccacaagg tttgctttgg cttgggacct catctgctgt catttagtaa
tgaataaaat 11520 tgctgacagt aataggatta actgtgtgtg gagatagcca
gggttagaga taaaaacact 11580 ggagaagtca aataagttgc tcgaggtcct
ctagctaata agctattaag tgggagagtg 11640 agggctagaa acaggccatc
tgtctcccaa gcacatgtcc attagtggtt tgctgatagc 11700 cttccagaac
aacagagagg actctcaaac atggtcttgc ctccctccaa ttgatcccct 11760
ccatgtgcct cacagcgggt ctttctaaaa ttaagttctg attttaattc tcccttgcta
11820 tagcacttag gtatggcttt cagccgtgca ataaaaagca ggcaagagtg
gctcaatcat 11880 ataggaggtt gtttttctta gatcccaagc aggtaatcct
gggcattatg gttgttctgc 11940 gtttatcaag gagccaaatt ctctatcacc
tcctgttcta tcctcctcag tatctggctc 12000 tattcttcag catctcaaga
tggcttgtgc tcctccaagc atggcagtca aattccacac 12060 aagaggggga
aatatgaagg gcagacagtg ctggtctcct gagctgtccc tctttgtcgg 12120
ggaaataaat gtattccttc aagtcccgtg agacttctga agtagacgtc tgcttacgtc
12180 tcacccacca gaactatgta aactgcacat agtgctaggt ctacatagcc
actcataact 12240 gccagggggt gggaaatctt taaataggtg taccaccaca
caattaggat gctaatagta 12300 agggagaagg agagaatagg ttttgcgcaa
gccaccagca tgcctgccac aattgcttaa 12360 aattcttcat tgacccctca
ttgccacagg atgaaatcca aacgccttct tagttgggaa 12420 tctgacctac
ctgtctctcc cacctggttc agacaccatt ctccttggtc ataaaattcc 12480
agtcatttgt gaacatccag ctcccccatg cctccatgcc tttgcacatg ctgttctttt
12540 atcttttatg ttgtcctttt atcttttatc caaaagagat atcccatcat
cacatctctt 12600 ttgtcagccc ccaaatactt tgtctttcaa gttcagctgg
aggattacct cctatttgaa 12660 atcagctttg tctcttacaa ccaaacaagg
ttttccttcc gagacactcc cacagcacct 12720 tgaactcatc tctatcaatc
attcatttga ttgtaatgaa gttgttggtg gtatgcctgt 12780 gtctctgaca
catctgcgat ctcatgagtt ccttaagtgg aatgtgaata gcgggatgaa 12840
cagtattggt cttcagccct catctctgca gatgttgctt gacccaaatg agcgttgcct
12900 tttattttga ttttgctttg atttgtctac tccatgtact tgagccatgc
atttctgtct 12960 tagcgatgct ttttaaaagt cattttttgg ttgattatcc
agatttgtcc acctttgctt 13020 ctagttgtag aaaaggatga agaaaatgga
gttttgcttc tagaactaaa tcctcctaac 13080 ccgtgggatt cagaacccag
atctcctgaa gatttggcat ttggggaagt gcaggtaagg 13140 aaatgttaaa
ttgcaatatt cttaaaaaca caaataaagc taacatatca atttatatat 13200
atatatatat atatattttt tttttttttt acatcttata ttaccttgag tattcttgga
13260 agtggctagt taggacatat aataaagtta ttctgaagtc tttttttttc
tttttccatg 13320 gtgagcagtg gcttgatgtg gatctcagct cccagacgag
gcactgaacc tgagccgcag 13380 tggtgaaagc accaagttct agccactaga
ccaccaggga actccctatt ctaaattctt 13440 gagcacatta tttaggaacc
tcaggaactt ggcaggatta caggaaatat atctagattt 13500 aaaaaaaaat
cttttaacag aggtcccaaa ggagagtcat gcacagctat gggaggaagt 13560
tcagaaactg cccttgctac cagatcactg tcagataaaa tggccagcta catgtttctg
13620 cacattgccc taagatcttt acaaactttt ctgtgcattt ttccactttt
aaaagaaaat 13680 ttcggggttc ctgttgttgc tcagtggtta acgaacccaa
ctagtatcca tggggacagg 13740 ggttcgagcc ctggcctcac tcagtgggtt
aagaatctgg cattgctgtg gctgtggcgt 13800 aggctggcgg ctacagctca
gattggaccc ctagcctgag aacctccata tgccgcaggt 13860 atggccctaa
aaaaaaaaaa aaagagagag agagaatttc ctccagaaaa aacactttgg 13920
tagtttggga gaagtaaaca accaaaaatt aatttttctg gagtattcgg gaagcttgta
13980 aaaatgggct cttacttttt tgaggagaca aatgggaacc tacccagaag
aggcacaatc 14040 acctgcattt gatttcttga cctctcccta ccttctttgc
tggctttcca catttggatt 14100 tctgtgacct tatctctgct ccttggtgtt
ttcatttttc ctgtggacgt gccagactat 14160 gggaagggag taaggcgttg
atttagaatc ctgtagtctc tgcctgtctc tagtcattgt 14220 tttcaccctt
ctcaaaggac cttgacatcc tgagtgagtc cgcaagtaat ttaggggaga 14280
agccttagaa gccagtgcag ccaggctaca tgactgtgtc cacccactgg aaccagtcat
14340 ttttatacct attcacagcc cccctaccat ttaaatcccc agaggtctgc
cataacatct 14400 gtaactccct ttcctggtaa attgtgttct aaaagactgg
taacaaaaga tattctgtgg 14460 tacagagcat aattaaatac ctgggagctg
atttgagtgg ggtaaatcaa ctggtttgac 14520 ccctaaaacc caccatgagc
atttctgttc taataaagta atgcccgtgc tgggaattgt 14580 gttctacgga
aatgctcctg ctgtgtcttt cttgagtcct gtgtcattga acatgcttag 14640
gagcaaaggt cccccatgtg gcttgtctgc taaccagccc agttccttgt tctggctggt
14700 aatgatccga tcatctgaat ctcactgtct tccaacagat cacgtacctt
actcacgcct 14760 gcatggacct caagctgggg gacaagagaa tggtgttcga
cccttggtta atcggtcctg 14820 cttttgcgcg aggatggtgg ttactacacg
agcctccatc tgattggctg gagaggctga 14880 gccgcgcaga cttaatttac
atcagtcaca tgcactcaga ccacctgag 14929 50 17221 DNA Porcine 50
aggaatggaa agcccaattc attaaaacag aaaggaagaa actcctgaac tacaaggctc
60 ggctggtgaa ggacctacaa cccagaattt actgcccctt tcctgggtat
ttcgtggaat 120 cccacccagc agacaagtat ggctggatat tttatataac
gtgtttacgc ataagttaat 180 atatgctgaa tgagtgattt agctgtgaaa
caacatgaaa tgagaaagaa tgattagtag 240 gggtctggag cttattttaa
caagcagcct gaaaacagag agtatgaata aaaaaaatta 300 aatacgtatg
gctggatatt ttatataacg tgtttacgca taagttaata tatgctgaat 360
gagtgattta gctgtgaaac aacatgaaat gagaaagaat gattagtagg ggtctggagc
420 ttattttaac aagcagcctg aaaacagaga gtatgaataa aaaaaattaa
atacaagagt 480 gtgctattac caattatgta taatagtctt gtacatctaa
cttcaattcc aatcactata 540 tgcttatact aaaaaacgaa gtatagagtc
aaccttcttt gactaacagc tcttccctag 600 tcagggacat tagctcaagt
atagtcttta tttttcctgg ggtaagaaaa gaaggattgg 660 gaagtaggaa
tgcaaagaaa taaaaaataa ttctgtcatt gttcaaataa gaatgtcatc 720
tgaaaataaa ctgccttaca tgggaatgct cttatttgtc aggtatatta aggaaacaaa
780 catcaaaaat gacccaaatg aactcaacaa tcttatcaag aagaattctg
aggtggtaac 840 ctggacccca agacctggag ccactcttga tctgggtagg
atgctaaagg acccaacaga 900 caggtttgac ttgaatattt acagggaaca
aaaatgattt ctgaattttt tcatgtttat 960 gagaaaataa agggcatacc
tatggcctct tggcaggtcc ctgtttgtag gaatattaag 1020 tttttcttga
ctagcatcct gagcttgtca tgcattaaga tctacacacc accctttaaa 1080
gtgggagtct tactgtataa aataaactat taaataagta tctttcaact ctggggtggg
1140 gggggagact gagttttttc acagtcctat ataataattt tcttatccta
taaaataatt 1200 aggagttccc gtagtggctc agcaatagca aacccgacta
gtatcgatga ggatgcgggt 1260 tcgattcctg gcccccctca gtgggttaag
gatctggcat tgccgtgagc tgtggtgtag 1320 gtggcagaca cggctcagat
cccacgttac tgtggctgtg gcataggcca gcagctccag 1380 ctctgattag
acccttagcc tgggaacttc catatgctgt gggtgtggcc ttgaaaaaaa 1440
ataaataaat aagataatta ctcaaatgtt ttccttgtct cagaacctta cttcaggata
1500 aagagtgaga aagttttttt tatgaagggc cattattaca gctcaaaaat
aagttgtctt 1560 cagcaagtag aaagcaataa gcctgagagt tagtgttcct
atcagtgtaa atattacctc 1620 ctcgccaatc cccagacagt ccatttgaac
aattaacggt gccctgggag tacagttcag 1680 aaacattaat gtggatgttc
cagacctgta tttttataag tacttgtctt gagccggatg 1740 gaaccatcat
tcctcaccat tatttagaag tggactgtga ctctgttgga gatcagggca 1800
cacggttacc aaaagcacac ccttctcctg gccttacctt tgcaaagctg gggtctggga
1860 cacagtcagc tgattatacc cttttactaa cttcccacag ctcaaatctg
gtcaattctc 1920 cttcacaaat ctcttaaaaa tccatcactc acctccagcc
tcttctgctg tggccttgat 1980 tcagcctctc acaatttttt tttaaccaga
attctggcag tggcccctga cttgcctctg 2040 tgctcccagc cccgctgtcc
tctgatccat cctccatgcc agcctttttc aatctgctgg 2100 tcacgattca
ttgatgggtt aggaaatcaa tggcatcaca actagcattt agaaaaagga 2160
aataggcgtt cccgccgtgg cacagcagaa ataaatccga ctaggaacca taaggttgcg
2220 ggttcaaccc ctggccttgt tcagtgggtt aaggatccgg cattgccgtg
ggctgttttg 2280 taagtcacag acatggctct gatccggcat tgctgtggct
ctggcgtagg cctgcagcat 2340 cagctccaat tagaccccta tcctgggagc
ctccatatgc tgcaagtgca gccctaaaaa 2400 aaataaaaaa ataaaaaaaa
ataaataaaa gaagtagaca aattgtatag aacaaccctg 2460 agtatgttgc
ctgagcacat ataacaaggg taagtattat ttcaggaaac tctggtttca 2520
cagatactct tggcatatgg acccctagag tcctgatgta aaatatattc ttcctgggat
2580 cttaggcaag aagtttgaaa gctccaactc tgcactgctg ccaaagaaat
gatttttaag 2640 tgcaaaactc ttcccgttcc cttccctgta taaaattcca
taggatctct ccagtgcctc 2700 taggataaag gcagttttca ttctctagtt
caaggtgaga gaagatttta attatttcac 2760 gttttagtgg ggaattcaag
agtctggcac ctgacatttg ctgaactctc tccattatcc 2820 ctctctagtt
ccccagacgc atcctatggt agaaattcgc aaactagagt gagcgtcaga 2880
gtaacccaag gaaactgggt aaatgcagct ccctgggctc taccccctga gattctgatt
2940 cagtagatct gaagcagagc cctggaatat gcatatgcat cattgtgtca
caccaagcat 3000 tctgggtaat gagagttgat gttaggttct cagtagtaag
acaagtatag agattccggg 3060 ggactgagtg ctcagctctg ccttggggag
gagggagagg gctaaagaga acaggagatg 3120 gggacaggga atgctcaacc
tccaatctta ggcatttgag ctatgtctta ggggtcagga 3180 ggaggttacc
aatatagtga ttaagagatt gaggttccag tcagagggat atgctggaga 3240
aggggggtga aaataatgtc ataggtttgg tgagtgcaga tactttgagt tttttaatat
3300 ttttattgaa atatagttga tttacaatgc tcttagtgag tacaattact
ttgaataagt 3360 gcatagatgt atgccattct tccagaaatg atttattgag
ctcctttggg catcatgcta 3420 agtacagggg aaacagctgt gaagaggtcc
ttcccttatg aagtcattca tccccttcag 3480 taaatgaagg taaaggaaaa
ggatgagaca gggacgccgt gttggaccag ggtcagaaag 3540 gccttataag
accttgcctg gagggcaagg aacttgcctg tgagtaagga gagcttgaga 3600
aagcgataaa gcaaagaagg aacattactg cattgtgttt tagaaaaacc atgtcctggg
3660 gaagaactcc tagagtcagg ggggccagtt gggagactgt gcttttttcc
aggaggagat 3720 aagtgaggct gctggctgag atggagcaag gatttagaga
agcagatatg agattcattt 3780 agaagttaga cattttagga tctgacacat
aatttatcac caaaaccagt gcatctctgg 3840 ctttgggcca ccagttttgg
agaagtggaa tgtagggacc taccattacc tgccaatctt 3900 tactacacag
atgcctattt ccctcctcat atttcctttc tccagatcac gtcctattct 3960
attgccagga ctcaagattc caccttgcat gcagtgatcc atcttcacac tggatggaca
4020 gctctaggga tgtcagagca cactcttgtc catactgctg actgggtctc
ctgtcagccc 4080 atctgtctat cagctgtggt attattagta taataagagg
gctgtatatg agagacacaa 4140 aattctaggt gtagctcaaa gataggctag
agttattcct atgtacaaca aatatttatg 4200 ggaccccttc tgtgtactgt
catggttgct gctttcatca tacttgtagt ctaatggagg 4260 tgggggcagg
gcaggaataa gcggatgtcc acaaaatcag taagaccact tatattcaac 4320
attttcataa tttagttatt tgagcccaaa gggtccacat ccgtggtatt ccaacttttt
4380 tttccccgga catggatctt tatctttttt tttttttctt ttttgcggcc
agacctgcgg 4440 catatggaag ttcccaggcc aggggttgaa tgggagttgc
agctgcctgg tctacaccac 4500 agccacagca aggtgggatc tgagctgcat
ctgtgacata caccgcagct gaggtaacac 4560 cagattctga acccactgaa
tgaggccagg gatggaaccc gtctccttat gaacactatg 4620 tcatgttctt
caccctctga gccacaacgg gaactccaga cttcgtcttt aaatgtattc 4680
tgacttggag agctatcaca ctaagcaatt aacaggagct gacctggttt aggctggggt
4740 ggggccctac tcctcaatgt tccctgaggc acatctgtgg gacccctggg
catcatctat 4800 ctgagcagcc ttagagctgc tcatccagtt gactgttgat
gtagaagtgc aaacttctgc 4860 cttccttatt tgttgctttc ttttttcatt
gttctctccc ctttgtgtct ttaagcaagg 4920 gcatcgtaga gcctccagaa
gggactaaga tttacaagga ttcctgggat tttggcccat 4980 atttgaatat
cttgaatgct gctataggag atgaaatatt tcgtcactca tcctggataa 5040
aagaatactt cacttgggct ggatttaagg attataacct ggtggtcagg gtatgctatg
5100 aagttattat ttgtttttgt tttcttgtat tacagagcta tatgaaaacc
tcttagtatt 5160 ccagttggtt tctcaataag cattcattga gccttactga
ctgtcagacg gagggcgtat 5220 tggactatgt gctgaaacaa tcctttgttg
aaaatgtagg gaatgttgaa aatgtaggga
5280 atgaaatgta gatccagctc tgtttctctt ttggaggatt ctttttcctc
catcaccgtg 5340 tcttggttct tgtttgtttt gggtttttgt gggtgttgta
ttgtgttgtg ttggttatgg 5400 cagtgacagc tatttaaact gtgaaacggg
ggagttcccg tcgtggcgca gtggttaacg 5460 aatccgactg ggaaccatga
ggttgcgggt tcggtccctg cccttgctca gtgggttaac 5520 gatccggcgt
tgccgtgagc tgtggtgtag gttgcagaca cggctcggat cccgcgttgc 5580
tgtggctcta gcgtaggcca gcggctacag ctccgattgg acccctagcc tgggaacctc
5640 catatgccgc aggagcggcc caaagaaata gcaaaaagac aaaataaata
aataaataaa 5700 taagtaagta aaataaactg tgaaacgggg agttcccttc
atggctcagc agttaacaaa 5760 cccagctagg atccatgagg atgtaggttc
gatccctggc cttgctcagt gggttaagaa 5820 tccagcgttg ctgtgagctg
tgatgtaggt cgcagatgca gcccagatcc tgcattgctg 5880 tggctgtggc
gtaggctggc agctgaagct ccgattcaac ccctagcctg ggaacatcca 5940
tatgctgcag gtgtggcctt aagaggcaaa aaaataaaaa aataaaaaat aaataaattg
6000 tgggacagac aggtggctcc actgcagagc tggtgtcctg tagcagcctg
gaagcaggta 6060 aggtaaggac tgcagctggg taaggactga attgcaccaa
ctgggaagta agcctagatc 6120 tagaacttaa gttagccctg acatagacac
acagagctca ccagctaagt ggttcagctt 6180 ataagctggt cactgaaact
gaggatgtcc acaaaagcaa aataagtagc aacaggcagc 6240 gggatgcaag
agaaagagga ggcctaaaat ggtctgggaa tccctgccat acctatattt 6300
tatcctactt atatttagtg cctgaatgtg tgcctggaga gcaaagttta gggaaagcat
6360 cgggaaatgc acagtattca tacccttagg aacaaagatc agttacctcc
agggtaaaga 6420 ctatttccaa gtttaaattt caacccctga acattagtac
tgggtaccag gcaacacttg 6480 ccatcctcaa aatcaatgaa tcctaaaatt
caacctgggg gtcagtgaca gtctgtgaca 6540 aagtttttgc tggtcagtaa
cgaaataagt atgagcacca tctgagtatg gtcaccaaga 6600 tgtcaactct
ctttcctttg gacgaattgt cattattcca agattaggtc ctttctattt 6660
ttgaggtgtg aaaacatctt tcctttcata aaataaaagg atagtaggtg gaagaatttt
6720 ttttgttttt tggtcttttt gctatttctt tgggccgctt ctgcagcata
tggaggttcc 6780 caggccaggg gtcgaatcgg agctttagcc accggcccac
gccagagcca cagcaacacg 6840 ggatccaagc cgcatctgca gcctacacca
cagctcacgg caatgccgga tcgttaaccc 6900 actgagcaag ggcagggacc
gaacccgcaa cctcatggtt cctagtcgga ttcgttaacc 6960 actgcgccac
gacgggaact cctaatgata ctcttttata tttagctact atgtgatgat 7020
gagaaacagt ccacatttta ttatttttta gccaatttga tatctcatta ctaagataat
7080 gataattttc tctataaatt ttatttaagt tagtgttatg aagtggtttt
gctagtgtag 7140 aaggctagga tttgaattca gttcaagaaa gaagagaggg
agggagggag agggatgggt 7200 agagggatgg ggcagtggga gagagcaaag
aggagagaca gtttttgtat taattctgct 7260 tcattgctat catttaaggg
cacttgggtc ttgcacattc tagaattttc taaggacctt 7320 gaccgccaga
ttgatatgct tcttcccttt accatgttgt catttgaaca gatgattgag 7380
acagatgagg acttcagccc tttgcctgga ggatatgact atttggttga ctttctggat
7440 ttatcctttc caaaagaaag accaagccgg gaacatccat atgaggaagt
aagcaggaat 7500 accagtggaa gtgccccttt cttccttcct tcctaaataa
acttttttat tttggaacaa 7560 ctttagagtt acagaaaagt tgcaaagata
ttatagacag tagtgtttat atatatatat 7620 aaattttttt ttgcttttta
tgaccacacc tgtggcatat ggaggttccc agtctagggg 7680 ttgaattgga
gctacagctg ccagtctgtg ccataaccac agcaatgcag gatctgggcc 7740
acgtctgtga cctacaccaa agctcacagc tggattctta acccactgag caaggccagg
7800 gattgaacct gcatcctcgt ggttcctagt tggattcgtt tccgctttgc
cgcaatggga 7860 actccaaatt attgttaata tcttacttta ctggggtaca
tttgttacaa ccaatactct 7920 gatactgaaa cattactgtt aactccgtac
ttgcttcttt ttgagtcatt tgcaaagact 7980 ggcttcttga cctgcttcct
tccaaacagc tggcctgcct atgctgttct cagacctgca 8040 agcactgatc
tctgcccccc ttgccttctc tccagtggtg tctccttccc caaacaaacc 8100
cagtgtggct ctggaaaggg agttaagtca acataaacca acacatattt tgttgagctc
8160 caattttgag caaatccctc acctacggca gacaggcatg atgttaagaa
ctagggcttt 8220 ggacacaagg tcaagaccaa gaagggttcc tcacccctac
tgattcagat aaccaataat 8280 gaggctttga atccctgtcc aaaggttgtt
ttttttccct tctattgagc ttcttgccac 8340 cttatcagtt ttttttatga
cagtcaaatg acatgatata tgtgagcata catggtaatt 8400 tttaattcta
tataaatgaa tcactaaata aattaggagg atatatagtc cacctttaag 8460
cgtattacac gtgtcacatg aatgtgtggc gacttaattg tagaggttta aatgtagctt
8520 cctataatag atgtgttcct aaactacatt ttaatcattg gacttgtatt
tttatgttag 8580 cacttgctgt tgaagaaaag cctatgccaa aagttcagtg
aaaccaataa tccactgcca 8640 gctttctgag ttaaaaaaaa tccctgggtt
ttcacacaca ggaacaccct gtgtgaaaca 8700 ctcatttaga gcaaaatgca
tctgataagg agttcctgtt gtgcctcaac tggttaagga 8760 cctgacattc
tccatgagaa tgtgagtttg atccccggcc ccactcgatg ggttaaggat 8820
ctggtgttgc cacaaactgc agctccgatt catctcctag cctagaaact tccacagccc
8880 agaatatgcc acagaattcg gctgtttaaa aaaaaaaaga aaaaaaaaag
aatcataaat 8940 gtgttggttt gttcaccaaa tacatgataa cttgctcttg
ccaagctcag cttcataaat 9000 attaagtcat ttaatacagc agccacctta
tgaacagata ttactatact tcccatttac 9060 agataaggaa aatgccatat
ttaaccaaga gattaaataa ctttcccgag gtcttatagc 9120 aagtaaatca
tggtgcaggg gtttgaccac acgcagtcta tctccagagt ctgtgtattt 9180
agccactgtt ttactttcaa atttaaattt ataaaacttc taaattatct gttaaccata
9240 atctttggaa tttttaaaac cacgagttcc tataaaatgt ttcattgaaa
gtaagtcact 9300 tttccatagc ttttgataat acatctgtag gataaagtaa
gccacagctc tcttgcagac 9360 ttggtacacc ctggggcaaa gcatcatgcc
tgtcacgtac atggtggtcc ttactttgac 9420 tctcagtgct tttattgccc
aggaattttg tgagatttct agttgttgag gtttgtttaa 9480 agaggttatg
ccggtacttg gaagagctct tttcttgcta cctggagcct tctcatattt 9540
cctttttgag gagggacatg aattgccttt caaactcata aatatatttt ctagtacaca
9600 agtctccatc ttccttagac gcatggctcc tggagttctc catcctcctg
ctccactttg 9660 ggtgggctcc tctctgggtc tgccaccaat ctgccaccca
gagacatcct tgacccactt 9720 ccagacccca ccatggcttc actttcttcg
ctttcctcct ttgtggaacc ttctgcttaa 9780 gaatctgagg aagaaaattt
gcacgtgagc taaactggag gtactttcct gcctggtctt 9840 gcacgatagc
ttggctgagc ccatgatgct gggtggctgt tactttccat ggacacccga 9900
aggcgttgct cctttggctt ctagttgcat gcagtgttgc ttatcccagg ctgatctttc
9960 ttccactgta ggtgactttt aagaattaag ggattaatct atatctacaa
caacaacaac 10020 aaagaccttt tcaagctgag gtagggcttt ctgtatatgt
ttggagtggt tatccagcag 10080 actttacttg aaggcagggg tcatatcctc
aagtgctcat aaacggacca cagaaagatc 10140 tcataattgg gtggagctgg
gtggggaccg tgtcatgtgg ccaggaaatg ccagatggga 10200 agggagtggc
ccttactgag ctccagctga actctgaatt ttctagaaaa ctcagaaatc 10260
tggatttttc atgtgtaata cccagattta tagatgtgga aagctaattc tttttttttt
10320 taagggacta taggcaatga actaagatct aggttgtatt tggacaaggg
gtcatcagtt 10380 taagctgtgt agttgagcgc tcagctattg ggctgaggga
cccctaaata ctgagacggg 10440 gaggtccttg ctctggggca tcacaagtac
actccctggt ctcattcaaa cacttttcct 10500 acaaaattga tcccatttct
tcagtgcact gtctgaatgc atttggccca gagccgtgct 10560 gaggcatagg
gaaggggtcc acggtttcat ggcatcgttt tgtgctgtgt gtccctgctg 10620
tcgtccagga tacctacctc tcctcctcct gcatctgaat gtccccccac agactctctg
10680 ggattctaca gcctctggcc tgttcctcag acacctctta cctgccagct
ttccagattc 10740 acattagtta gtccaaatct actgccgtca gtgactcact
tcatttcttc ttctccgagg 10800 cagttcagcc cggtacagtt gttttgtcaa
cacttcagtt gagtctggaa gatgtgcatg 10860 ggttatgcac gagagcggtc
catcattttg agctagaagt cctttctcag cccagagaca 10920 agtcctcatc
tcctttactt cctgactctt cttcctctgc atccttccaa gatatctctt 10980
tctccagcca ccacctaaat ctcttctttt cccggggttc cgtgctcaac ccactcttct
11040 tcttaaatct gtggctgggt gaacgcatct gctggcacca cttctctgct
aaagactcca 11100 aaaatccata ggtcctgccc ggcctttgcc cacctctctc
caacactgtc cagctttaga 11160 tgtagagcta atccccccag agatatcatt
ccctggatgt ctaagtcctt tggtatctca 11220 ctttcagcgt gttcaaaatc
ctcttacaac tgttctttct ccttttccat cttgattatt 11280 ggcaacatgc
cagcctttcc cctaccccca gcagtgagcc aagctagaaa caagggctta 11340
atcttcaatc tttccttctc catccctaaa cctaatgagt ctccaagccc ttcccagttt
11400 acaccctaaa tgttgctcaa aacatcccct agttcttcca cgtgctctcc
tctatattga 11460 aaggtcaaga aaggccatct tccctccact gtgaggaaat
agatcttgat actgcccctg 11520 agctgggcag tcctcgacct gacaaactgt
gcagtgtttc taaatctcta ctggcaaaat 11580 gagagtgcct ttgacctgtg
ttgcgatctc agatcacagt ggatgtaatt gttttatagg 11640 aatggtgaac
gaaaaagaag taaatcccta atgccaaact cctgatcatt ctatgtcatt 11700
taatagcctg tcatttatga taaagtttcc tctactggca ttagcacaat acttctcagg
11760 aaaaaaaaat atgatgccag atactgaaaa gctcctgggt aaacatgaac
atgggtaccg 11820 ataaaatggt gaagccagtc caatcttaga gtgacttccc
ttcatgctac ttcatgctct 11880 tttttttttt tttttttaag aaaaacccct
tttttttttc tcacaccagt cacagaggag 11940 accgaggctt agcaaggtta
aggtcacatg attagtaagt gctgggctga aactcaaaac 12000 catctctgct
tgtctcctaa ccctgtgcac ctctgactat tcaacagatc ctgtgtcagg 12060
agttgggatt ctttgaaggt aagggccttg accaccgaat taaggtaatc ttgctctgtg
12120 gcaggccttg ttttcagtat tttaagtaca ctggctcagg taatcctcac
aacagcccca 12180 ggaggaatgt tctattacct ccactgtata gatgaggaac
ttgaggcaca gaatggttgc 12240 caaggtcaca cagctatatt gggggttcat
acccagccat ccaactctgt ctgtactctc 12300 tgccactctg cacccccagc
tcctgatcca cttcctgttt ccatccctcg atttctgctg 12360 cactcagggg
cccctctccc cctcggcctg tgagatctgc ttcagtaggc ttttctccct 12420
gactcctcca tccctgtcct tacaggcagc tgcttctctc cgggacacga ggggtccata
12480 cggacactct ctactggctg ggttgcgcct aactcgtgat tcctcctctg
tttcagattc 12540 ggagccgggt tgatgtcatc agacacgtgg taaagaatgg
tctgctctgg gatgacttgt 12600 acataggatt ccaaacccgg cttcagcggg
atcctgatat ataccatcat ctgtaagtcc 12660 gaaaatgcct gtcgtgtgtg
ccttaggctg ctgcggagga ggccagggct atataagcag 12720 agtcagtgac
tgactgtgcc ctgcagtgtt gatggccatg gagattccac cgttagagct 12780
tttttctttg ttaaccttga aggcaaatct ggttaggaag ataactttca aagagtcacc
12840 atctggacat tcatgcccat gtgcttcaat cctgtataca agcagtttag
agtacaggga 12900 agggaaggac attatgaaag ggagagggtg tgtttggatc
cagcagctcc atcctcagaa 12960 tttatctgaa gacactgcaa aattactaag
aatcactatg acaagaatga ggatggggtg 13020 atatggcaaa gttgtgatcc
tggaagacct tcatctccca tgttgcccaa ctctgaacat 13080 gaatttggtg
aactagttgg ttaaggggat gatcctccaa gtttctccct ggttgagctc 13140
caaaaaccat gtaagtttct catagcaaaa ccgtataggt ccttagggct ttagttggaa
13200 tatttgtgct gaaatgctgg aaagccccat ttgccatttt tgtatttgca
aaataatcat 13260 caagagggga gaatgcattc tttcatgacc actgaccctc
tgaaaaggtc aggaatttag 13320 tctgaagtag gcaagcctcc taccccgctt
ctgccatgag cttgcacgca caggcctgtc 13380 ttgacatttc ttctttatag
atttcttttt gaatatcttg aaattgcttt aaaaatattt 13440 aaagaatgta
gaattatata aaataaaaag gaaataaccc cacacctccc acaaaaccct 13500
gtttcctgcc tttctccacc cactctccag ggtaacactt ggtaacagca tagttgtatc
13560 accccaggcc tatttttgag catatcagca tttcaagaaa tgtatttttt
ctcaataaaa 13620 catcccttat agttgaggag gggaggttat cattcctggg
ttttgttttt tttttttttt 13680 taatgtaatc ctggtacatc ggtaatttgc
attttttatt cattaatatc tttggtattt 13740 ctagtgttgg gacacacagg
tcaacctcag tttttgggtt tttttttttg tctttttgtc 13800 tttctagggc
cacacctgca gcatatggac gttcccaagc taggagtcta atcagagctg 13860
tagccaccag cctacgtcat agccatagca acgtcagatc caagccgtgt ctgtgaccta
13920 caagcacagc tcatggcaac accggatcct taaccactga acgaggccag
gggatcgaac 13980 acacatcctc atggatccta gtcatgttca ttaaccactg
agtcatgatg ggaactccaa 14040 cttcaactat tttaatgtct gtaaaacatt
ccatttggaa accatttcat ttgtaaagca 14100 aaatgaaaac attttgttca
ttttcaacag agttcgtagc tgacttctgt tctggaaaaa 14160 aggaaatgga
gcaaatttga gtgagaaaga ttcaaagata acttttcttt taaaaaaaat 14220
tatatcttgg aaacttctgg gctattgatt ctgaagacta tttttctata tactgttttg
14280 atagcaaagt tcataaatgt gaaaggatcc tgcgatgaat cttgggaagc
agtcatagcc 14340 caatatatct ttgttgcttt taaaatgaga tttagtttac
taaatatttt tctgatcata 14400 aaaataacac agatctaccg cagaaaattt
ggaaaaaaaa aaacttttaa attcaaaaaa 14460 cagttaaacc acaaatgatc
ccaccatcca gagagcaatt tgtactttgg tgtctagttc 14520 atctttcttt
ttctgtttac aagcacatat accacaagca ttttttcaaa aaatgaaaat 14580
gggataatac tatacatacg tctgtacacc tgcatagtta ctgaacagtc tttgatctac
14640 cctgtaagtt tctaactttt cattatttga aatgatgttt tggcaaagaa
atatgtaggt 14700 gtgtctcgca cactttcata atgatttctt aggataaatt
tcttaggata aattcataat 14760 gatttcttat aataatccat actctgccaa
ctgatcttca gggaagccaa ctcgccttct 14820 cagaaataac atataaccca
tttacttgcc ctctcaccaa tactaggtcc taatgttttt 14880 gtgtacagat
tctatatttt tacatacaag aattccttaa agcaaggcat gtcacagaaa 14940
aatagaagga agacacaatt gtcatgttta aggactgcat tctgtaccaa aaatgctaag
15000 ttaaatgaac atctgaaaca gtacagaaac gctatctttc agggaaagct
gagtaccagg 15060 tactgaacag attttggcaa atacagcagg catggatgtt
tccaaaacat gtttttctac 15120 tttatctctt acaggttttg gaatcatttt
caaataaaac tccccctcac accacctgac 15180 tggaagtcct tcctgatgtg
ctctgggtag agaggacctg agctgtccca ggtaaagcat 15240 cctgcaggtc
tgggagacac tcttattctc cagcccatca cactgtgttt ggcatcagaa 15300
ttaagcaggc actatgccta tcagaaaacc tgacttttgg gggaatgaaa gaagctaaca
15360 ttacaagaat gtctgtgttt aaaaataagt caataaggga gttcccatcg
tggctcagtg 15420 gtaacgaacc ctactagtat ccattgagga cacaggttca
atatctggcc tcactcagtc 15480 ggctaaggat ccagtgatgc cgtgagctgc
agtgtaggcc acagacgtgg ctcagatctg 15540 gtgctgctgt ggctatggtg
taggccggcc ccctgtaact ccaattcgac ccctaggctg 15600 ggaacctaaa
aagaccccaa aaaagtcgct ttaatgaata gtgaatacat ccagcccaaa 15660
gtccacagac tctttggtct ggttgtggca aacatacagc cagttaacaa acaagacaaa
15720 aattatccta ggtggtcagt gggggttcag agctgaatcc tgaacactgg
aaggaaaaca 15780 gcaaccaaat ccaaatactg tatggttttg cttatatgta
gaatctaaat tcaaagcaaa 15840 tgagcaaacc aattgaaaca gttatggaag
acaagcaggt ggttgtcagg ggggagataa 15900 ggggaggcag gaaagacctg
ggcgagggag attaagaggt accaactttc agttgcaaaa 15960 caaatgagtc
accagtatga aatgtgcaat gtgggaaata caggccataa ctttataatc 16020
tctttttttt ttttgtcttt tttgcctttt ctaaggctgc tcccgtggca tatggaggtt
16080 cccaggctag gagtccaaac agagctgtag ctgccagcct acaccagagc
cacagcaaca 16140 cgggaacctt aacccgctga gcaaggccag ggatcgaacc
cgagtcctca cagatgccag 16200 tagggttcat taaccactga gccacgacag
gaattccagg gtctgttgtg ttcttaaaac 16260 acttccagga gagtgagtgg
tatgtcataa gtaaacaata aatgttaacc acaacaagct 16320 tatgaaataa
acaggaaagc catatgacct acaatcagtc attgggagaa tccacaaaag 16380
gttgagcaga ggatcaattc cagctcacac tccagtttta gattctcccc tgccttaaag
16440 catcacagac tacataatct gagctgaaga ataaaaatta aaactcaccc
cagtgcaaaa 16500 cagaaatgaa aaagtattaa aacgaggttc atactgttgt
tcattagcaa tatcttttat 16560 tcacaggggt gcccaacaac atgaaaaaat
caagaattta ttgctgctac gtcaaagctt 16620 ataccagaga ttatgcctta
tagacattag caatggataa ttatatgttg cacttgtgaa 16680 atgtgcacat
atcctgttta tgaatcacca catagccaga ttatcaatat tttacttatt 16740
tcgtaaaaaa tccacaattt tccataacag aatcaacgtg tgcaatagga acaagattgc
16800 tatggaaaac gagggtaaca ggaggagata ttaatccaag catagaagaa
atagacaaat 16860 gaggggccat aaggggaata tagggaagag aaaaaaatta
agatggaatt ttaaaaggag 16920 aatgtaaaaa atagatattt gttccttaat
aggttgattc ctcaaataga gcccatgaat 16980 ataatcaaat aggaagggtt
catgactgtt ttcaattttt caaaaagctt tgttgaaatc 17040 atagacttgc
aaaacaaggc tgtagaggcc accctaaaat ggaaaatttc actgggactg 17100
aaattatttt gattcaatga caaaatttgt tatttactgc ggattataaa ctctaacaaa
17160 tagcgatctc tttgcttcat aaaaacataa acactagcta gtaataaaat
gagttctgca 17220 g 17221 51 26 DNA artificial sequence Primer 51
catggacctc aagctggggg acaaga 26 52 28 DNA artificial sequence
primer 52 gtgttcgacc cttggttaat cggtcctg 28 53 28 DNA artificial
sequence Primer 53 caggaccgat taaccaaggg tcgaacac 28 54 26 DNA
artificial sequence primer 54 tcttgtcccc cagcttgagg tccatg 26 55 27
DNA artificial sequence primer 55 ctcctggaag cttctgtcaa gacgaac 27
56 27 DNA artificial sequence primer 56 gcctgataca cagtgctgtg
caatggt 27 57 29 DNA artificial sequence primer 57 accacccaag
tctggaatct tcttacact 29 58 30 DNA artificial sequence primer 58
gactctcata caaaagctaa gctgggtaag 30 59 30 DNA artificial sequence
primer 59 gactctcata caaaacctaa gctgggtaag 30 60 30 DNA artificial
sequence primer 60 gactctcata caaaacctag gctgggtaag 30 61 30 DNA
artificial sequence primer 61 gactctcata caaaacctag gctaggtaag 30
62 28 DNA artificial sequence primer 62 ccttatactg gccccaattg
gatcttac 28 63 28 DNA artificial sequence primer 63 ccttatactg
gccccaattg gatcttac 28 64 30 DNA artificial sequence primer 64
cttacctagc ctaggttttg tatgagagtc 30 65 28 DNA artificial sequence
primer 65 gacaaaccac aattggaatg cactcgag 28
* * * * *
References